{"gene":"FOXO1","run_date":"2026-04-28T17:46:04","timeline":{"discoveries":[{"year":1999,"finding":"Akt (protein kinase B) directly phosphorylates FOXO1 (FKHR) in vitro and in vivo at three sites (Thr-24, Ser-256, Ser-319), leading to suppression of FOXO1-mediated transcriptional activation; mutation of all three sites to alanine renders FOXO1 resistant to Akt inhibition and causes apoptosis in a DNA-binding-dependent manner.","method":"In vitro kinase assay, co-transfection with active/inactive Akt, alanine-substitution mutagenesis, reporter gene assays, apoptosis assay","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 1 — in vitro phosphorylation plus mutagenesis, independently replicated in two back-to-back papers (PMIDs 10358014 and 10358075)","pmids":["10358014","10358075"],"is_preprint":false},{"year":1999,"finding":"FOXO1 binds in vitro to the insulin-responsive sequence (IRS) in the IGFBP-1 promoter and activates transcription from IRS-containing reporter plasmids; Akt-mediated phosphorylation inhibits this transcriptional activity.","method":"In vitro DNA binding assay, luciferase reporter assay, co-transfection","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 1 — direct DNA-binding assay plus functional reporter, replicated across two concurrent papers","pmids":["10358014","10358075"],"is_preprint":false},{"year":2001,"finding":"FOXO1 confers insulin sensitivity onto glucose-6-phosphatase (G6P) expression in kidney epithelial cells; adenoviral transduction of FOXO1 restores insulin-mediated suppression of dexamethasone/cAMP-induced G6P expression, and dominant-negative FOXO1 partially inhibits G6P and PEPCK expression in primary hepatocytes, establishing FOXO1 as a mediator of insulin regulation of gluconeogenic gene expression.","method":"Adenoviral transduction, dominant-negative mutant expression, primary hepatocyte cultures, gene expression analysis","journal":"The Journal of clinical investigation","confidence":"High","confidence_rationale":"Tier 2 — loss-of-function and gain-of-function with defined molecular phenotype in multiple cell types","pmids":["11696581"],"is_preprint":false},{"year":2001,"finding":"DYRK1A phosphorylates FOXO1 (FKHR) specifically at Ser-329 in vitro; this phosphorylation reduces FOXO1 nuclear abundance and transcriptional activity; DYRK1A and FOXO1 co-localize in discrete nuclear regions and can be co-immunoprecipitated, suggesting DYRK1A phosphorylates FOXO1 at Ser-329 in vivo.","method":"Purification of kinase from skeletal muscle, in vitro kinase assay, co-immunoprecipitation, nuclear co-localization, site-directed mutagenesis, reporter assay","journal":"The Biochemical journal","confidence":"High","confidence_rationale":"Tier 1-2 — purified kinase reconstitution, co-IP, and localization data in one study","pmids":["11311120"],"is_preprint":false},{"year":2002,"finding":"FOXO1-mediated inhibition of cell cycle progression and transformation requires transcriptional repression of D-type cyclins (Class III targets) rather than activation of insulin-response-sequence genes; chromatin immunoprecipitation confirmed direct FOXO1 occupancy at cyclin D promoters.","method":"Transcriptional profiling, chromatin immunoprecipitation (ChIP), functional cell-cycle and transformation assays","journal":"Cancer cell","confidence":"High","confidence_rationale":"Tier 2 — ChIP plus functional epistasis with multiple orthogonal methods in one study","pmids":["12150827"],"is_preprint":false},{"year":2003,"finding":"Insulin-induced Akt-mediated phosphorylation of FOXO1 targets it for ubiquitination and proteasomal degradation; efficient ubiquitination requires both phosphorylation at the PKB consensus sites and cytoplasmic retention, establishing a two-step negative regulation (nuclear exclusion then proteasomal degradation).","method":"Proteasome inhibitor treatment, in vivo and in vitro ubiquitination assays, PI3K pathway inhibitors, phosphorylation-site mutants","journal":"Proceedings of the National Academy of Sciences of the United States of America","confidence":"High","confidence_rationale":"Tier 1-2 — in vitro ubiquitination reconstitution plus in vivo confirmation with multiple mutants","pmids":["13679577"],"is_preprint":false},{"year":2003,"finding":"The androgen receptor (AR) directly interacts with FOXO1 through protein-protein interaction (carboxyl terminus of FOXO1, aa 350–655, with AR A/B region and ligand-binding domain), blocking FOXO1 DNA-binding activity and its ability to induce Fas ligand expression and apoptosis in an AKT-independent manner.","method":"In vivo and in vitro co-precipitation, domain-mapping, reporter gene assay, apoptosis/cell-cycle assays","journal":"Molecular and cellular biology","confidence":"High","confidence_rationale":"Tier 2 — reciprocal co-precipitation with domain mapping and functional consequence","pmids":["12482965"],"is_preprint":false},{"year":2004,"finding":"CK1 phosphorylates FOXO1a at Ser-322 and Ser-325 within a multisite phosphorylation domain in vivo; pharmacological inhibition of CK1 with D4476 specifically blocks these phosphorylations and attenuates IGF-1/insulin-induced nuclear exclusion of FOXO1a.","method":"Cell-permeable CK1 inhibitor (D4476), phospho-specific analysis, nuclear/cytoplasmic fractionation, H4IIE hepatoma cells","journal":"EMBO reports","confidence":"High","confidence_rationale":"Tier 2 — site-specific pharmacological inhibition with subcellular localization readout","pmids":["14710188"],"is_preprint":false},{"year":2004,"finding":"Foxo1 is essential for normal vascular development; Foxo1-deficient mice die at embryonic day ~11 with severe defects in angiogenesis, and Foxo1-null endothelial cells derived from ES cells show aberrant morphological response to VEGF, indicating Foxo1 is required for proper endothelial cell response to VEGF signaling.","method":"Foxo1 knockout mice (embryonic lethal phenotype), ES-cell-derived endothelial cell differentiation assay, VEGF stimulation","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 2 — constitutive knockout with defined vascular phenotype and in vitro mechanistic follow-up","pmids":["15184386"],"is_preprint":false},{"year":2004,"finding":"SIRT1 (Sir2) binds and deacetylates FOXO1 at lysine residues (K242, K245, K262) that are acetylated by CBP, reversing the attenuation of FOXO1 transcriptional activity; SIRT1 is recruited to FOXO1 target promoters (MnSOD, p27kip1) and enhances their expression in a deacetylase-activity-dependent manner.","method":"Co-immunoprecipitation, in vitro deacetylation assay, ChIP, reporter assay, deacetylase-inactive mutants","journal":"Proceedings of the National Academy of Sciences of the United States of America","confidence":"High","confidence_rationale":"Tier 1-2 — biochemical reconstitution of deacetylation, ChIP, and catalytic mutants in one study","pmids":["15220471"],"is_preprint":false},{"year":2004,"finding":"Foxo transcription factors (including Foxo1) drive atrogin-1/MAFbx ubiquitin ligase expression in skeletal muscle; constitutively active Foxo3 acts on the atrogin-1 promoter to cause transcription and dramatic muscle atrophy; dominant-negative Foxo or RNAi-mediated knockdown prevents atrogin-1 induction and muscle atrophy in vitro and in vivo.","method":"Constitutively active Foxo constructs, dominant-negative constructs, RNAi in mouse muscle in vivo, promoter reporter assay, myotube atrophy assay","journal":"Cell","confidence":"High","confidence_rationale":"Tier 2 — genetic epistasis with multiple loss- and gain-of-function approaches in vitro and in vivo","pmids":["15109499"],"is_preprint":false},{"year":2005,"finding":"FoxO1 protects pancreatic beta cells against oxidative stress by forming a complex with PML and SIRT1 to activate NeuroD and MafA transcription; acetylation of FoxO1 targets it to PML and prevents ubiquitin-dependent degradation, and constitutively nuclear FoxO1 in beta cells prevents hyperglycemia-induced suppression of MafA.","method":"Co-immunoprecipitation (FoxO1-PML-SIRT1 complex), acetylation-defective and acetylation-mimicking mutants, transgenic mice, in vivo MafA expression analysis","journal":"Cell metabolism","confidence":"High","confidence_rationale":"Tier 2 — complex reconstitution by co-IP, functional mutagenesis, and in vivo transgenic validation","pmids":["16154098"],"is_preprint":false},{"year":2005,"finding":"Acetylation of FOXO1 by CBP at Lys-242, Lys-245, and Lys-262 reduces its DNA-binding affinity by neutralizing positive charges required for DNA interaction; additionally, acetylation increases susceptibility of FOXO1 to Akt-mediated phosphorylation at Ser-253, and DNA binding inhibits PKB phosphorylation of FOXO1 in vitro.","method":"In vitro DNA binding assays, in vitro kinase assays, acetylation-mimicking and acetylation-deficient mutants, phosphorylation analysis","journal":"Proceedings of the National Academy of Sciences of the United States of America","confidence":"High","confidence_rationale":"Tier 1 — in vitro biochemical reconstitution of acetylation-DNA binding crosstalk with mutagenesis","pmids":["16076959"],"is_preprint":false},{"year":2005,"finding":"Skp2, an oncogenic F-box protein, interacts with and ubiquitinates FOXO1 in an Akt phosphorylation (Ser-256)-dependent manner, promoting FOXO1 proteasomal degradation and abolishing its growth-inhibitory and pro-apoptotic functions.","method":"Co-immunoprecipitation, in vitro ubiquitination assay, phosphorylation-site mutants, cell proliferation/survival assays, mouse lymphoma model","journal":"Proceedings of the National Academy of Sciences of the United States of America","confidence":"High","confidence_rationale":"Tier 1-2 — biochemical reconstitution of ubiquitination with site-specific mutants and in vivo model","pmids":["15668399"],"is_preprint":false},{"year":2005,"finding":"Foxo1 (and Foxo3a) overexpression inhibits endothelial cell migration and tube formation; Foxo1 specifically represses angiopoietin-2 expression and binds to the eNOS promoter to repress eNOS expression, establishing Foxo1 as a regulator of angiogenesis-related gene expression in endothelial cells.","method":"Constitutively active Foxo constructs, siRNA knockdown, gene expression profiling, ChIP on eNOS promoter, migration/tube formation assays","journal":"The Journal of clinical investigation","confidence":"High","confidence_rationale":"Tier 2 — ChIP plus gain/loss-of-function with defined molecular and cellular phenotypes","pmids":["16100571"],"is_preprint":false},{"year":2005,"finding":"PAK1 (p21-activated kinase 1) directly interacts with FOXO1 (FKHR) and phosphorylates it in response to estrogen stimulation, causing perinuclear cytoplasmic retention and preventing FOXO1 from activating Fas ligand promoter transcription; dominant-negative PAK1 restores FOXO1 transcriptional activity.","method":"Co-immunoprecipitation, in vitro kinase assay (PAK1 phosphorylating FKHR), localization microscopy, reporter gene assay","journal":"FEBS letters","confidence":"Medium","confidence_rationale":"Tier 2 — direct kinase assay and co-IP, single lab","pmids":["12560069"],"is_preprint":false},{"year":2005,"finding":"Beta-catenin directly binds FOXO transcription factors and enhances FOXO transcriptional activity in mammalian cells; this interaction is evolutionarily conserved (C. elegans BAR-1/DAF-16) and is enhanced under oxidative stress, establishing beta-catenin as a co-activator of FOXO-dependent stress response.","method":"Co-immunoprecipitation, reporter assay, C. elegans genetic epistasis, oxidative stress experiments","journal":"Science","confidence":"High","confidence_rationale":"Tier 2 — direct binding demonstrated by co-IP, replicated across mammalian cells and C. elegans","pmids":["15905404"],"is_preprint":false},{"year":2005,"finding":"FoxO1a directly activates transcription of the cyclic GMP-dependent protein kinase I (cGKI) gene; the resulting cGKI then phosphorylates FoxO1a and abolishes its DNA-binding activity, establishing an autoregulatory feedback loop that controls myoblast fusion.","method":"Reporter gene assay, phosphorylation assay (cGKI on FoxO1a), myoblast fusion assay","journal":"Molecular and cellular biology","confidence":"Medium","confidence_rationale":"Tier 2 — direct kinase assay plus functional consequence in differentiation, single lab","pmids":["16107711"],"is_preprint":false},{"year":2006,"finding":"Constitutively active FoxO1 in transgenic mouse liver increases gluconeogenic gene expression (PEPCK, G6Pase), suppresses glycolytic, pentose phosphate shunt, and lipogenic genes (including glucokinase and SREBP-1c), and impairs glucose tolerance; adenoviral FoxO1 in hepatocytes confirms direct regulation of these metabolic gene programs.","method":"Transgenic mice with liver-specific constitutively active FoxO1, gene array, adenoviral transduction of hepatocytes, metabolic phenotyping","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 2 — in vivo transgenic and in vitro adenoviral models with genome-wide gene expression analysis","pmids":["16492665"],"is_preprint":false},{"year":2006,"finding":"FOXO1A cooperates with HoxA-10 to regulate decidual IGFBP-1 expression; co-immunoprecipitation and ChIP demonstrate physical association of FOXO1A with HoxA-11 and binding of both to the decidual prolactin promoter enhancer, forming a decidual-specific transcriptional complex that switches HoxA-11 from repressor to activator.","method":"Co-immunoprecipitation, ChIP, reporter gene assay, siRNA knockdown","journal":"PloS one","confidence":"Medium","confidence_rationale":"Tier 2 — co-IP and ChIP with functional reporter assay, single lab","pmids":["19727442"],"is_preprint":false},{"year":2008,"finding":"PRMT1 methylates FOXO1 at Arg-248 and Arg-250 within the Akt phosphorylation consensus motif; this arginine methylation directly blocks Akt-mediated phosphorylation of FOXO1 at Ser-253 in vitro and in vivo, preventing nuclear exclusion and proteasomal degradation, thereby promoting oxidative-stress-induced apoptosis.","method":"In vitro methylation assay, in vitro kinase assay, PRMT1 siRNA knockdown, polyubiquitination assay, apoptosis assay, phosphorylation-deficient mutants","journal":"Molecular cell","confidence":"High","confidence_rationale":"Tier 1 — in vitro biochemical reconstitution of methylation-phosphorylation crosstalk with mutagenesis, multiple orthogonal methods","pmids":["18951090"],"is_preprint":false},{"year":2008,"finding":"PGC-1alpha cooperates with FoxO1a and HNF-4alpha to drive selenoprotein P (SeP) promoter activity; PGC-1alpha enhances FoxO1a-stimulated SeP transcription through a FoxO-responsive element; insulin suppresses this axis, establishing FoxO1a as a component of the PGC-1alpha-regulated selenium homeostasis network.","method":"Reporter assay, promoter mutagenesis, adenoviral overexpression, primary rat hepatocytes, mRNA/protein analysis","journal":"Hepatology","confidence":"Medium","confidence_rationale":"Tier 2 — promoter mutagenesis and overexpression in primary cells, single lab","pmids":["18972406"],"is_preprint":false},{"year":2010,"finding":"FoxO1 promotes inflammation by enhancing TLR4-mediated signaling in macrophages; ChIP-Seq shows FoxO1 binds multiple enhancer-like elements within the Tlr4 gene and TLR4 pathway genes; TLR4 activation induces Akt which inactivates FoxO1, establishing a self-limiting inflammatory feedback loop.","method":"ChIP combined with massively parallel sequencing (ChIP-Seq), FoxO1 knockout macrophages, reporter assay, inflammatory cytokine measurements","journal":"The EMBO journal","confidence":"High","confidence_rationale":"Tier 2 — genome-wide ChIP-Seq plus loss-of-function in primary macrophages, multiple orthogonal methods","pmids":["21045807"],"is_preprint":false},{"year":2010,"finding":"TAK1-NLK (Nemo-like kinase) pathway phosphorylates FOXO1 at Pro-directed Ser/Thr residues in the transactivation domain; this phosphorylation inhibits FOXO1 transcriptional activity and causes nuclear exclusion independently of PI3K/Akt; NLK binds FOXO1 and this interaction is evolutionarily conserved in Drosophila.","method":"Co-immunoprecipitation, kinase assay (NLK on FOXO1), nuclear/cytoplasmic fractionation, siRNA knockdown of TAK1/NLK, FOXO target gene mRNA assay, PARP cleavage","journal":"The Journal of biological chemistry","confidence":"Medium","confidence_rationale":"Tier 2 — direct binding and kinase assay with functional localization data, single lab","pmids":["20061393"],"is_preprint":false},{"year":2010,"finding":"Cytosolic FoxO1 mediates autophagy independently of its transcriptional activity; in response to oxidative stress or serum starvation, FoxO1 is acetylated by dissociation from SIRT2, and acetylated FoxO1 binds to Atg7 (an E1-like autophagy protein) to induce autophagy and cell death; this is associated with tumor suppressor activity.","method":"Co-immunoprecipitation (FoxO1-Atg7 interaction), transcription-deficient FoxO1 mutants, autophagy assays, SIRT2 interaction studies, xenograft tumor model","journal":"Nature cell biology","confidence":"High","confidence_rationale":"Tier 2 — direct protein-protein interaction by co-IP, functional mutants separating transcriptional from cytoplasmic role, in vivo tumor model","pmids":["20543840"],"is_preprint":false},{"year":2011,"finding":"FoxO1 and SIRT1 regulate beta-cell responses to nitric oxide; FoxO1 translocates from cytoplasm to nucleus in response to nitric oxide and stimulates GADD45α expression for DNA repair; SIRT1 regulates this process—SIRT1 inhibition shifts FoxO1 from a protective (GADD45α/DNA repair) to a proapoptotic (PUMA/caspase-3) program.","method":"Nuclear translocation assay, gene expression analysis, SIRT1 pharmacological inhibitors, siRNA, caspase-3 cleavage assay","journal":"The Journal of biological chemistry","confidence":"Medium","confidence_rationale":"Tier 2 — direct localization experiments with functional consequence in primary beta cells, single lab","pmids":["21196578"],"is_preprint":false},{"year":2011,"finding":"Hepatic deletion of Foxo1 (but not Foxo3 or Foxo4 alone) reduces blood glucose by decreasing gluconeogenic enzyme gene expression; combined deletion of Foxo1 and Foxo3 additionally increases hepatic lipid synthesis and secretion, demonstrating that Foxo1 specifically controls gluconeogenesis while Foxo3 regulates lipogenic gene expression.","method":"Liver-specific Cre/LoxP conditional knockout mice (individual and combined deletions), metabolic phenotyping, liver transcript analysis","journal":"Endocrinology","confidence":"High","confidence_rationale":"Tier 2 — conditional knockout with genetic epistasis distinguishing paralog-specific functions in vivo","pmids":["22147007"],"is_preprint":false},{"year":2013,"finding":"mTORC2 promotes inactivating phosphorylation of class IIa HDACs, leading to acetylation of FoxO1 (and FoxO3), which releases c-Myc from a suppressive miR-34c network to drive glycolytic reprogramming in glioblastoma; this is an Akt-independent mechanism linking mTORC2 to FoxO acetylation and cancer metabolism.","method":"mTORC2 genetic manipulation, HDAC phosphorylation/acetylation assays, FoxO acetylation analysis, c-Myc/miR-34c network analysis, glioblastoma patient samples","journal":"Cell metabolism","confidence":"High","confidence_rationale":"Tier 2 — mechanistic epistasis across multiple nodes with clinical correlation, multiple orthogonal methods","pmids":["24140020"],"is_preprint":false},{"year":2014,"finding":"FOXO1/3 knockdown in vascular smooth muscle cells phenocopies PTEN deficiency by increasing Runx2 protein via decreased Runx2 ubiquitination, promoting vascular calcification; AKT activation causes FOXO1/3 nuclear exclusion, establishing a PTEN/AKT/FOXO1/3/Runx2 ubiquitination axis controlling VSMC calcification.","method":"SMC-specific PTEN knockout mice (Cre/LoxP), siRNA knockdown, in vitro calcification assay, Runx2 ubiquitination assay, ex vivo arterial calcification","journal":"Arteriosclerosis, thrombosis, and vascular biology","confidence":"Medium","confidence_rationale":"Tier 2 — in vivo genetic model plus in vitro mechanistic dissection, single lab","pmids":["25378413"],"is_preprint":false},{"year":2015,"finding":"FOXO1 directly regulates CCR7 and ICAM-1 promoters (shown by ChIP and promoter reporter assays) in dendritic cells; FOXO1 nuclear localization is induced by bacteria via the MAPK pathway; lineage-specific FOXO1 deletion impairs DC phagocytosis, chemotaxis, and lymph node homing, which can be rescued by ICAM-1 and CCR7 re-expression.","method":"ChIP, promoter reporter assay, lineage-specific knockout (CD11c.Cre), rescue with plasmid transfection, in vivo lymph node homing assay","journal":"Journal of immunology","confidence":"High","confidence_rationale":"Tier 2 — ChIP plus genetic rescue in vivo demonstrating direct target gene mechanism","pmids":["25786691"],"is_preprint":false},{"year":2015,"finding":"FOXO1 enhances keratinocyte migration and wound healing by upregulating TGFβ1 under normal glucose conditions; under high glucose, FOXO1 nuclear localization is induced but FOXO1 does not bind the TGFβ1 promoter—instead it drives SERPINB2 and CCL20 expression that inhibits migration; insulin treatment or FOXO1 silencing rescues impaired migration in high glucose.","method":"Lineage-specific Foxo1 deletion, FOXO1 promoter binding analysis, siRNA, high-glucose cell culture, wound healing migration assay","journal":"The Journal of cell biology","confidence":"High","confidence_rationale":"Tier 2 — in vivo conditional knockout with mechanistic promoter binding and rescue experiments","pmids":["25918228"],"is_preprint":false},{"year":2016,"finding":"FOXO1 suppresses MYC signaling in endothelial cells to restrain glycolysis and mitochondrial respiration, acting as a gatekeeper of endothelial quiescence; endothelial-restricted FOXO1 deletion causes increased EC proliferation and vessel hyperplasia, while forced FOXO1 expression restricts vascular expansion; restoration of MYC in FOXO1-overexpressing endothelium normalizes metabolism and branching.","method":"Endothelial-specific knockout and overexpression mouse models, metabolic flux assays (glycolysis, respiration), MYC rescue experiments, EC proliferation assays","journal":"Nature","confidence":"High","confidence_rationale":"Tier 2 — in vivo endothelial-specific genetic models with mechanistic epistasis and metabolic phenotyping, multiple orthogonal approaches","pmids":["26735015"],"is_preprint":false},{"year":2017,"finding":"DDB1-CUL4A E3 ubiquitin ligase promotes FOXO1-driven hepatic gluconeogenesis by degrading CRY1 (a known DDB1 target); loss of Ddb1 in hepatocytes downregulates FOXO1 protein and impairs gluconeogenic response; CRY1 depletion increases nuclear FOXO1 abundance and enhances gluconeogenesis, linking circadian clock protein degradation to FOXO1 stability.","method":"Hepatocyte-specific Ddb1 knockout mice, Cry1 depletion in vivo, FOXO1 protein stability analysis, gluconeogenic gene expression, glucose production assay","journal":"Diabetes","confidence":"Medium","confidence_rationale":"Tier 2 — conditional knockout with mechanistic pathway dissection, single lab","pmids":["28790135"],"is_preprint":false},{"year":2017,"finding":"SCP4 (CTDSPL2) is a nuclear phosphatase that directly dephosphorylates FoxO1/3a, promoting nuclear retention and transcriptional activation of PEPCK1 and G6PC gluconeogenic genes; SCP4 overexpression increases hepatic glucose production; SCP4 gene ablation causes neonatal hypoglycemia.","method":"In vitro dephosphorylation assay (SCP4 on FoxO1/3a), ectopic expression, siRNA knockdown, SCP4 knockout mice (neonatal hypoglycemia), glucose production assay","journal":"Diabetes","confidence":"High","confidence_rationale":"Tier 1-2 — direct in vitro dephosphorylation reconstitution plus in vivo knockout phenotype","pmids":["28851713"],"is_preprint":false},{"year":2018,"finding":"FOXO1 directly regulates VEGFA expression in keratinocytes (shown by in vitro promoter assay); keratinocyte-specific FOXO1 deletion reduces VEGFA expression in wounds, resulting in reduced endothelial cell proliferation, reduced angiogenesis, and impaired wound re-epithelialization.","method":"Keratinocyte-specific Foxo1 knockout mice, VEGFA promoter reporter assay, endothelial proliferation assay, porcine wound model with FOXO1 inhibitor","journal":"The Journal of pathology","confidence":"High","confidence_rationale":"Tier 2 — in vivo conditional knockout with direct promoter binding evidence and large-animal model","pmids":["29574902"],"is_preprint":false},{"year":2018,"finding":"Foxo1 directly binds the Il9 and Irf4 promoters in Th9 cells (shown by ChIP and luciferase assays) to transactivate them; pharmacological or genetic disruption of Foxo1 in CD4+ T cells reduces IL-9 and promotes IL-17A/IFNγ; adoptive transfer of Foxo1-inhibitor-treated Th9 cells reduces asthma-like symptoms.","method":"ChIP, luciferase promoter assay, Foxo1 pharmacological inhibitor (AS1842856), CD4-specific Foxo1 knockout, adoptive transfer airway allergy model","journal":"Scientific reports","confidence":"Medium","confidence_rationale":"Tier 2 — ChIP plus loss-of-function with defined cellular phenotype, single lab","pmids":["29339772"],"is_preprint":false},{"year":2019,"finding":"Glucagon suppresses hepatic mitochondrial complex III and IV activity and reduces mitochondrial biogenesis (NRF1, TFAM, MFN2 expression) in a FOXO1-dependent manner; hepatocyte-specific Foxo1 deletion largely rescues glucagon-induced mitochondrial dysfunction, establishing FOXO1 as a mediator of glucagon's effects on mitochondrial function.","method":"Foxo1 hepatocyte-specific knockout mice, glucagon treatment, mitochondrial function assays (complex activity), db/db diabetic mice","journal":"The Journal of endocrinology","confidence":"Medium","confidence_rationale":"Tier 2 — conditional knockout with functional mitochondrial phenotype, single lab","pmids":["31026811"],"is_preprint":false},{"year":2020,"finding":"METTL14 promotes FOXO1 expression in endothelial cells by directly binding FOXO1 mRNA, increasing its m6A modification, and enhancing translation through YTHDF1 recognition; METTL14 also interacts with FOXO1 protein and together they act on VCAM-1 and ICAM-1 promoters to promote inflammatory gene transcription.","method":"Methylated RNA immunoprecipitation (RIP) sequencing, RIP assay, co-immunoprecipitation, ChIP, METTL14 knockout mice, TNF-α inflammation model","journal":"Theranostics","confidence":"Medium","confidence_rationale":"Tier 2 — RIP-seq, co-IP, and ChIP with in vivo knockout, single lab","pmids":["32802173"],"is_preprint":false},{"year":2020,"finding":"FOXO1 binds directly to the KLF5 promoter in cardiomyocytes to increase KLF5 expression; KLF5 in turn binds the NOX4 promoter to induce NOX4 expression, reactive oxygen species, and ceramide accumulation, causing diabetic cardiomyopathy; cardiomyocyte-specific FOXO1 deletion protects from diabetic cardiomyopathy by reducing KLF5.","method":"ChIP (FOXO1 on KLF5 promoter; KLF5 on NOX4 promoter), cardiomyocyte-specific FOXO1 knockout mice, AAV-mediated KLF5 delivery, pharmacological KLF5 inhibition, oxidative stress and ceramide measurements","journal":"Circulation research","confidence":"High","confidence_rationale":"Tier 2 — ChIP-validated direct target gene regulation, in vivo knockout, and multiple orthogonal rescue approaches","pmids":["33539225"],"is_preprint":false},{"year":2020,"finding":"S100A11 interacts with HDAC6 to inhibit HDAC6 deacetylase activity, thereby releasing FOXO1 from HDAC6 binding and increasing FOXO1 acetylation; acetylated FOXO1 activates autophagy and lipogenesis programs promoting liver steatosis.","method":"Co-immunoprecipitation (S100A11-HDAC6 interaction), HDAC6 inhibitor, FOXO1 inhibitor, adenoviral overexpression, in vivo high-fat diet model, lipid staining","journal":"Cellular and molecular gastroenterology and hepatology","confidence":"Medium","confidence_rationale":"Tier 2 — co-IP with pharmacological and genetic dissection in vitro and in vivo, single lab","pmids":["33075563"],"is_preprint":false},{"year":2021,"finding":"Akt phosphorylation of FoxO1 is required for Sox10 expression in oligodendrocytes; unphosphorylated (nuclear) FoxO1 represses the Sox10 promoter; Akt-triple-knockout oligodendrocyte lineage mice lack mature oligodendrocytes and myelination with downregulated Sox10, identifying a FoxO1 phosphorylation-dependent mechanism for oligodendrocyte differentiation.","method":"OL lineage-specific Akt1/2/3 triple conditional knockout mice, Sox10 promoter reporter assay, FoxO1 phosphorylation-site mutants, ChIP (FoxO1 on Sox10 promoter)","journal":"The Journal of neuroscience","confidence":"Medium","confidence_rationale":"Tier 2 — in vivo triple knockout with direct promoter occupancy evidence, single lab","pmids":["34385359"],"is_preprint":false},{"year":2021,"finding":"Foxo1 in intestinal epithelial cells regulates goblet cell autophagy and mucus secretion; IEC-specific Foxo1 deletion impairs goblet cell autophagy and mucus secretion, leading to gut microbiota dysbiosis, disrupted tight junction organization, and increased susceptibility to intestinal inflammation.","method":"Intestinal epithelial cell-specific Foxo1 conditional knockout mice, goblet cell autophagy assays, microbiome analysis, tight junction protein analysis, colitis susceptibility assay","journal":"The Journal of experimental medicine","confidence":"Medium","confidence_rationale":"Tier 2 — in vivo conditional knockout with defined cellular mechanism (autophagy-mucus axis), single lab","pmids":["34287641"],"is_preprint":false},{"year":2022,"finding":"FoxO1 in hepatic macrophages (Kupffer cells) promotes pro-inflammatory M1 polarization by counteracting Stat6 signaling; myeloid-specific FoxO1 knockout skews macrophage polarization from M1 to M2 and reduces hepatic inflammation, steatosis, and fibrosis in NASH.","method":"Myeloid-specific FoxO1 conditional knockout (LysM-Cre), M1/M2 polarization markers, Stat6 pathway analysis, NASH diet model, high-fat diet model","journal":"The Journal of clinical investigation","confidence":"High","confidence_rationale":"Tier 2 — in vivo conditional knockout with mechanistic epistasis (FoxO1-Stat6) and disease-relevant phenotype","pmids":["35700043"],"is_preprint":false},{"year":2023,"finding":"FOXO1 directly binds the promoter regions of CD36 and STEAP4 genes in bovine adipocytes (shown by ChIP and site-directed mutagenesis) and regulates their transcriptional activity, linking FOXO1 to lipogenesis through the AMPK and PI3K/AKT pathways.","method":"ChIP, site-directed mutagenesis of FOXO1 binding sites, transcriptomic and metabolomics analyses, siRNA knockdown","journal":"International journal of biological macromolecules","confidence":"Medium","confidence_rationale":"Tier 2 — ChIP plus promoter mutagenesis, single lab, non-human model","pmids":["37506793"],"is_preprint":false}],"current_model":"FOXO1 is a forkhead-family transcription factor that is negatively regulated by Akt-mediated phosphorylation (at Thr-24, Ser-256, Ser-319), which drives its nuclear exclusion, ubiquitination by Skp2, and proteasomal degradation; its activity is further modulated by CK1- and DYRK1A-mediated phosphorylation, CBP-mediated acetylation (attenuating DNA binding and increasing Akt sensitivity), SIRT1/SIRT2-mediated deacetylation (enhancing transcriptional activity or autophagy), PRMT1-mediated arginine methylation (blocking Akt phosphorylation), and dephosphorylation by SCP4; in the nucleus, FOXO1 binds insulin-response sequences to activate gluconeogenic genes (G6Pase, PEPCK) and pro-apoptotic targets (FasL, PUMA, GADD45α), represses D-type cyclins and MYC to enforce cell-cycle arrest and metabolic quiescence, and forms functional complexes with PML-SIRT1, beta-catenin, Smad proteins, HoxA factors, and Atg7 (in a transcription-independent autophagy pathway) to integrate insulin/PI3K, oxidative stress, TGFβ, and Wnt signals across diverse cell types including hepatocytes, endothelial cells, macrophages, beta cells, and T cells."},"narrative":{"teleology":[{"year":1999,"claim":"Identification of Akt as the direct kinase for FOXO1 at three conserved sites established the core mechanism by which insulin/PI3K signaling inactivates FOXO1 transcriptional activity and triggers apoptosis when this regulation is lost.","evidence":"In vitro kinase assay with purified Akt, alanine-substitution mutagenesis, reporter and apoptosis assays in mammalian cells","pmids":["10358014","10358075"],"confidence":"High","gaps":["Structural basis of Akt–FOXO1 interaction not resolved","In vivo phosphorylation kinetics not determined"]},{"year":1999,"claim":"Demonstration that FOXO1 binds insulin-response sequences and activates IRS-containing promoters defined it as a transcription factor directly mediating insulin-responsive gene regulation.","evidence":"In vitro DNA-binding assay and luciferase reporter in mammalian cells","pmids":["10358014","10358075"],"confidence":"High","gaps":["Genome-wide binding profile not yet mapped","Endogenous target genes not identified at this stage"]},{"year":2001,"claim":"Gain- and loss-of-function experiments in hepatocytes established FOXO1 as the mediator of insulin regulation of gluconeogenic genes (G6Pase, PEPCK), connecting FOXO1 phosphorylation to whole-body glucose homeostasis.","evidence":"Adenoviral FOXO1 transduction and dominant-negative mutant in primary hepatocytes and kidney epithelial cells","pmids":["11696581"],"confidence":"High","gaps":["Liver-specific in vivo deletion not yet performed","Relative contribution of FOXO1 versus other FOXO paralogs in hepatic gluconeogenesis unknown"]},{"year":2001,"claim":"Discovery that DYRK1A phosphorylates FOXO1 at Ser-329 to promote nuclear exclusion revealed a PI3K-independent input controlling FOXO1 activity.","evidence":"Purified kinase from skeletal muscle, co-IP, nuclear co-localization, site-directed mutagenesis","pmids":["11311120"],"confidence":"High","gaps":["Physiological contexts requiring DYRK1A–FOXO1 regulation not defined","In vivo significance not validated with DYRK1A knockout"]},{"year":2002,"claim":"ChIP-validated occupancy of cyclin D promoters showed FOXO1 enforces cell-cycle arrest through transcriptional repression of D-type cyclins rather than solely through gene activation, broadening the mechanistic repertoire.","evidence":"Transcriptional profiling, ChIP, cell-cycle and transformation assays","pmids":["12150827"],"confidence":"High","gaps":["Mechanism of transcriptional repression (corepressor identity) not determined"]},{"year":2003,"claim":"Reconstitution of phosphorylation-dependent ubiquitination and proteasomal degradation of FOXO1 established a two-step inactivation model (nuclear exclusion then degradation) downstream of insulin/Akt, with Skp2 identified as the E3 ligase.","evidence":"In vitro and in vivo ubiquitination assays with proteasome inhibitors and phospho-site mutants; Skp2 co-IP and in vitro ubiquitination","pmids":["13679577","15668399"],"confidence":"High","gaps":["Whether additional E3 ligases target FOXO1 in specific tissues not resolved","Deubiquitinases for FOXO1 not identified"]},{"year":2004,"claim":"CK1-mediated phosphorylation at Ser-322/325 was shown to cooperate with Akt phosphorylation to drive nuclear exclusion, revealing multisite phosphorylation as a combinatorial regulatory code.","evidence":"CK1 inhibitor D4476, phospho-specific analysis, nuclear/cytoplasmic fractionation in hepatoma cells","pmids":["14710188"],"confidence":"High","gaps":["CK1 isoform specificity not fully determined","Interplay with DYRK1A and NLK phosphorylation not tested"]},{"year":2004,"claim":"Foxo1-null embryonic lethality at ~E11 with angiogenic defects established FOXO1 as essential for vascular development and endothelial VEGF responses.","evidence":"Constitutive Foxo1 knockout mice, ES-cell-derived endothelial differentiation assay","pmids":["15184386"],"confidence":"High","gaps":["Downstream transcriptional targets mediating the vascular phenotype not fully defined at this stage"]},{"year":2004,"claim":"Demonstration that SIRT1 deacetylates FOXO1 at CBP-acetylated lysines and is recruited to FOXO1 target promoters established the acetylation/deacetylation switch as a major regulatory axis modulating FOXO1 transcriptional output.","evidence":"In vitro deacetylation with purified SIRT1, ChIP on MnSOD/p27 promoters, catalytic-dead mutant control","pmids":["15220471"],"confidence":"High","gaps":["Context-dependent selection of FOXO1 targets by acetylation state not systematically mapped"]},{"year":2005,"claim":"Biochemical reconstitution showed CBP acetylation at Lys-242/245/262 attenuates FOXO1 DNA binding by charge neutralization and sensitizes FOXO1 to Akt phosphorylation, revealing bidirectional crosstalk between acetylation and phosphorylation.","evidence":"In vitro DNA binding and kinase assays with acetylation-mimicking and acetylation-deficient mutants","pmids":["16076959"],"confidence":"High","gaps":["In vivo stoichiometry of acetylated versus unacetylated FOXO1 unknown"]},{"year":2005,"claim":"Discovery that FOXO1 forms a complex with PML and SIRT1 in pancreatic beta cells to activate NeuroD/MafA transcription under oxidative stress revealed a tissue-specific protective function coupling acetylation to PML body localization.","evidence":"Co-IP of FoxO1–PML–SIRT1 complex, acetylation mutants, transgenic mice with beta-cell phenotyping","pmids":["16154098"],"confidence":"High","gaps":["Whether PML–SIRT1–FOXO1 complex operates in non-beta-cell contexts not tested"]},{"year":2005,"claim":"Beta-catenin was identified as a direct FOXO co-activator under oxidative stress, conserved from C. elegans to mammals, linking Wnt pathway components to FOXO-dependent stress responses.","evidence":"Co-IP, reporter assay, C. elegans genetic epistasis (BAR-1/DAF-16)","pmids":["15905404"],"confidence":"High","gaps":["Competition between TCF/LEF and FOXO for beta-catenin binding not quantitatively resolved"]},{"year":2006,"claim":"Liver-specific constitutively active FOXO1 transgenics demonstrated that FOXO1 globally coordinates hepatic metabolism by activating gluconeogenesis while suppressing glycolytic, lipogenic, and pentose phosphate pathway genes.","evidence":"Transgenic mice with liver-specific active FoxO1, gene array, adenoviral hepatocyte transduction, metabolic phenotyping","pmids":["16492665"],"confidence":"High","gaps":["Direct versus indirect transcriptional targets within suppressed metabolic programs not distinguished"]},{"year":2008,"claim":"PRMT1-mediated methylation at Arg-248/250 was shown to directly block Akt phosphorylation at Ser-253, establishing arginine methylation as a third post-translational code that antagonizes the Akt-ubiquitination axis.","evidence":"In vitro methylation and kinase assays, PRMT1 siRNA, polyubiquitination assay, apoptosis assay","pmids":["18951090"],"confidence":"High","gaps":["Physiological stimuli triggering PRMT1-dependent FOXO1 methylation not defined","Demethylase not identified"]},{"year":2010,"claim":"ChIP-Seq in macrophages revealed FOXO1 occupancy at enhancer-like elements within TLR4 and downstream innate immune genes, with TLR4 activation feeding back through Akt to inactivate FOXO1—establishing a self-limiting inflammatory circuit.","evidence":"ChIP-Seq, FoxO1 knockout macrophages, reporter assays, cytokine measurements","pmids":["21045807"],"confidence":"High","gaps":["Precise chromatin remodeling events at FOXO1-bound enhancers not characterized"]},{"year":2010,"claim":"Cytoplasmic acetylated FOXO1 was shown to bind Atg7 and induce autophagy independently of transcription, revealing a non-nuclear tumor-suppressive function mediated by SIRT2 dissociation.","evidence":"Co-IP (FoxO1–Atg7), transcription-deficient mutants, autophagy assays, SIRT2 interaction studies, xenograft tumor model","pmids":["20543840"],"confidence":"High","gaps":["Structural basis of acetylated FOXO1–Atg7 interaction unknown","Relative contribution of transcriptional versus cytoplasmic autophagy pathways in different tissues not quantified"]},{"year":2011,"claim":"Liver-specific Foxo1 deletion confirmed its non-redundant role in gluconeogenesis in vivo, while combined Foxo1/Foxo3 deletion additionally derepressed lipogenesis, genetically separating paralog-specific metabolic functions.","evidence":"Cre/LoxP conditional single and combined knockouts, metabolic phenotyping, hepatic transcript analysis","pmids":["22147007"],"confidence":"High","gaps":["Mechanism by which Foxo3 specifically suppresses lipogenic genes independently of Foxo1 not fully delineated"]},{"year":2016,"claim":"Endothelial-specific genetic models demonstrated FOXO1 enforces vascular quiescence by repressing MYC, restraining glycolysis and proliferation; MYC re-expression rescued the FOXO1-overexpression phenotype, defining a FOXO1–MYC metabolic antagonism in angiogenesis.","evidence":"Endothelial-specific knockout and overexpression mice, metabolic flux assays, MYC epistasis rescue","pmids":["26735015"],"confidence":"High","gaps":["Whether FOXO1 directly represses MYC transcription or acts post-transcriptionally not resolved"]},{"year":2017,"claim":"Identification of SCP4 as the phosphatase that directly dephosphorylates FOXO1 to retain it in the nucleus provided the missing counter-regulatory enzyme to Akt; SCP4-null mice exhibit neonatal hypoglycemia, validating the axis in vivo.","evidence":"In vitro dephosphorylation assay, SCP4 knockout mice, hepatic glucose production assays","pmids":["28851713"],"confidence":"High","gaps":["Regulation of SCP4 itself under fasting/feeding transitions not characterized"]},{"year":2020,"claim":"FOXO1 was shown to directly bind the KLF5 promoter in cardiomyocytes, initiating a KLF5→NOX4→ROS→ceramide cascade underlying diabetic cardiomyopathy; cardiomyocyte-specific FOXO1 deletion was protective.","evidence":"ChIP (FOXO1 on KLF5 promoter), cardiomyocyte-specific knockout, AAV rescue, oxidative stress measurements","pmids":["33539225"],"confidence":"High","gaps":["Whether FOXO1-KLF5 axis operates in non-diabetic cardiac stress unknown"]},{"year":2022,"claim":"Myeloid-specific FOXO1 deletion revealed its role in promoting M1 macrophage polarization by opposing Stat6 signaling, connecting FOXO1 to inflammatory hepatic pathology in NASH.","evidence":"Myeloid-specific Foxo1 conditional knockout (LysM-Cre), M1/M2 markers, Stat6 pathway analysis, NASH/HFD models","pmids":["35700043"],"confidence":"High","gaps":["Molecular mechanism of FOXO1–Stat6 antagonism (direct binding vs. transcriptional) not defined"]},{"year":null,"claim":"Key unresolved questions include how tissue-specific cofactor assemblies select distinct FOXO1 transcriptional programs, the structural basis for the acetylated-FOXO1–Atg7 autophagy complex, the identity of FOXO1 demethylases and additional deubiquitinases, and how the multiple kinase inputs (Akt, CK1, DYRK1A, NLK, PAK1, cGKI, mTORC2/HDAC) are integrated in real time at the single-cell level.","evidence":"","pmids":[],"confidence":"Low","gaps":["No high-resolution structure of full-length FOXO1 with cofactors","Integrative quantitative modeling of multisite post-translational modification crosstalk lacking","Tissue-specific FOXO1 interactome not systematically mapped"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0140110","term_label":"transcription regulator activity","supporting_discovery_ids":[1,2,4,9,11,14,18,22,29,31,35,38]},{"term_id":"GO:0003677","term_label":"DNA binding","supporting_discovery_ids":[1,4,12,22,29,35,38,43]}],"localization":[{"term_id":"GO:0005634","term_label":"nucleus","supporting_discovery_ids":[0,3,7,9,11,12,20,25,33]},{"term_id":"GO:0005829","term_label":"cytosol","supporting_discovery_ids":[5,7,15,24]}],"pathway":[{"term_id":"R-HSA-74160","term_label":"Gene expression (Transcription)","supporting_discovery_ids":[1,2,4,18,22,29,31,38]},{"term_id":"R-HSA-1430728","term_label":"Metabolism","supporting_discovery_ids":[2,18,26,36,43]},{"term_id":"R-HSA-162582","term_label":"Signal Transduction","supporting_discovery_ids":[0,5,7,13,20,27,33]},{"term_id":"R-HSA-5357801","term_label":"Programmed Cell Death","supporting_discovery_ids":[0,6,13,20,25]},{"term_id":"R-HSA-9612973","term_label":"Autophagy","supporting_discovery_ids":[24,39,41]},{"term_id":"R-HSA-1640170","term_label":"Cell Cycle","supporting_discovery_ids":[4]},{"term_id":"R-HSA-168256","term_label":"Immune System","supporting_discovery_ids":[22,29,35,42]},{"term_id":"R-HSA-392499","term_label":"Metabolism of proteins","supporting_discovery_ids":[5,13,20]}],"complexes":["PML-SIRT1-FOXO1","FOXO1-Atg7","FOXO1-beta-catenin"],"partners":["AKT1","SIRT1","SIRT2","SKP2","PRMT1","CTNNB1","ATG7","PML"],"other_free_text":[]},"mechanistic_narrative":"FOXO1 is a forkhead-family transcription factor that serves as a central integrator of insulin/PI3K/Akt signaling with metabolic gene regulation, cell-cycle control, apoptosis, autophagy, and inflammatory programs across diverse cell types. Akt directly phosphorylates FOXO1 at Thr-24, Ser-256, and Ser-319, triggering nuclear exclusion, Skp2-mediated ubiquitination, and proteasomal degradation; additional kinases (CK1, DYRK1A, NLK, PAK1) and the phosphatase SCP4 fine-tune its subcellular localization, while CBP-mediated acetylation attenuates DNA binding and SIRT1/SIRT2-mediated deacetylation restores transcriptional activity or redirects FOXO1 to a cytoplasmic Atg7-dependent autophagy pathway, and PRMT1-mediated arginine methylation antagonizes Akt phosphorylation [PMID:10358014, PMID:13679577, PMID:15668399, PMID:14710188, PMID:15220471, PMID:16076959, PMID:18951090, PMID:20543840, PMID:28851713]. In the nucleus, FOXO1 binds insulin-response elements to activate gluconeogenic genes (G6Pase, PEPCK), pro-apoptotic targets (FasL, PUMA, GADD45α), and immune-regulatory genes (TLR4, CCR7, IL-9), while repressing D-type cyclins, MYC, and eNOS to enforce cell-cycle arrest, endothelial quiescence, and metabolic restraint [PMID:11696581, PMID:12150827, PMID:26735015, PMID:21045807, PMID:25786691, PMID:16100571]. Foxo1-null mice die at ~E11 from angiogenic defects, and tissue-specific deletions in liver, endothelium, macrophages, intestinal epithelium, and immune cells demonstrate non-redundant roles in gluconeogenesis, vascular remodeling, inflammatory polarization, goblet cell autophagy, and T-cell differentiation [PMID:15184386, PMID:22147007, PMID:35700043, PMID:34287641, PMID:29339772]."},"prefetch_data":{"uniprot":{"accession":"Q12778","full_name":"Forkhead box protein O1","aliases":["Forkhead box protein O1A","Forkhead in rhabdomyosarcoma"],"length_aa":655,"mass_kda":69.7,"function":"Transcription factor that is the main target of insulin signaling and regulates metabolic homeostasis in response to oxidative stress (PubMed:10358076, PubMed:12228231, PubMed:15220471, PubMed:15890677, PubMed:18356527, PubMed:19221179, PubMed:20543840, PubMed:21245099). Binds to the insulin response element (IRE) with consensus sequence 5'-TT[G/A]TTTTG-3' and the related Daf-16 family binding element (DBE) with consensus sequence 5'-TT[G/A]TTTAC-3' (PubMed:10358076). Activity suppressed by insulin (PubMed:10358076). Main regulator of redox balance and osteoblast numbers and controls bone mass (By similarity). Orchestrates the endocrine function of the skeleton in regulating glucose metabolism (By similarity). Also acts as a key regulator of chondrogenic commitment of skeletal progenitor cells in response to lipid availability: when lipids levels are low, translocates to the nucleus and promotes expression of SOX9, which induces chondrogenic commitment and suppresses fatty acid oxidation (By similarity). Acts synergistically with ATF4 to suppress osteocalcin/BGLAP activity, increasing glucose levels and triggering glucose intolerance and insulin insensitivity (By similarity). Also suppresses the transcriptional activity of RUNX2, an upstream activator of osteocalcin/BGLAP (By similarity). Acts as an inhibitor of glucose sensing in pancreatic beta cells by acting as a transcription repressor and suppressing expression of PDX1 (By similarity). In hepatocytes, promotes gluconeogenesis by acting together with PPARGC1A and CEBPA to activate the expression of genes such as IGFBP1, G6PC1 and PCK1 (By similarity). Also promotes gluconeogenesis by directly promoting expression of PPARGC1A and G6PC1 (PubMed:17024043). Important regulator of cell death acting downstream of CDK1, PKB/AKT1 and STK4/MST1 (PubMed:18356527, PubMed:19221179). Promotes neural cell death (PubMed:18356527). Mediates insulin action on adipose tissue (By similarity). Regulates the expression of adipogenic genes such as PPARG during preadipocyte differentiation and, adipocyte size and adipose tissue-specific gene expression in response to excessive calorie intake (By similarity). Regulates the transcriptional activity of GADD45A and repair of nitric oxide-damaged DNA in beta-cells (By similarity). Required for the autophagic cell death induction in response to starvation or oxidative stress in a transcription-independent manner (PubMed:20543840). Mediates the function of MLIP in cardiomyocytes hypertrophy and cardiac remodeling (By similarity). Positive regulator of apoptosis in cardiac smooth muscle cells as a result of its transcriptional activation of pro-apoptotic genes (PubMed:19483080). Regulates endothelial cell (EC) viability and apoptosis in a PPIA/CYPA-dependent manner via transcription of CCL2 and BCL2L11 which are involved in EC chemotaxis and apoptosis (PubMed:31063815)","subcellular_location":"Cytoplasm; Nucleus","url":"https://www.uniprot.org/uniprotkb/Q12778/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/FOXO1","classification":"Not Classified","n_dependent_lines":36,"n_total_lines":1208,"dependency_fraction":0.029801324503311258},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[{"gene":"DIAPH1","stoichiometry":0.2}],"url":"https://opencell.sf.czbiohub.org/search/FOXO1","total_profiled":1310},"omim":[{"mim_id":"621355","title":"KRI1 HOMOLOG; KRI1","url":"https://www.omim.org/entry/621355"},{"mim_id":"621330","title":"OXIDATIVE STRESS-RESPONSIVE PROTEIN, SERINE-RICH, 1; OSER1","url":"https://www.omim.org/entry/621330"},{"mim_id":"621297","title":"LYMPHATIC ENDOTHELIAL TRANSCRIPTIONAL REGULATOR lncRNA 1; LETR1","url":"https://www.omim.org/entry/621297"},{"mim_id":"621053","title":"NEDD4-BINDING PROTEIN 2-LIKE 1; N4BP2L1","url":"https://www.omim.org/entry/621053"},{"mim_id":"619008","title":"LONG INTERGENIC NONCODING RNA 598; LINC00598","url":"https://www.omim.org/entry/619008"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"Supported","locations":[{"location":"Nucleoplasm","reliability":"Supported"},{"location":"Cytosol","reliability":"Additional"}],"tissue_specificity":"Tissue enhanced","tissue_distribution":"Detected in all","driving_tissues":[{"tissue":"skeletal muscle","ntpm":84.4}],"url":"https://www.proteinatlas.org/search/FOXO1"},"hgnc":{"alias_symbol":["FKH1"],"prev_symbol":["FKHR","FOXO1A"]},"alphafold":{"accession":"Q12778","domains":[{"cath_id":"1.10.10.10","chopping":"164-232","consensus_level":"high","plddt":91.2925,"start":164,"end":232}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/Q12778","model_url":"https://alphafold.ebi.ac.uk/files/AF-Q12778-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-Q12778-F1-predicted_aligned_error_v6.png","plddt_mean":50.97},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=FOXO1","jax_strain_url":"https://www.jax.org/strain/search?query=FOXO1"},"sequence":{"accession":"Q12778","fasta_url":"https://rest.uniprot.org/uniprotkb/Q12778.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/Q12778/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/Q12778"}},"corpus_meta":[{"pmid":"10358014","id":"PMC_10358014","title":"Negative regulation of the forkhead transcription factor FKHR by Akt.","date":"1999","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/10358014","citation_count":654,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"10358075","id":"PMC_10358075","title":"Phosphorylation of the transcription factor forkhead family member FKHR by protein kinase B.","date":"1999","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/10358075","citation_count":604,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"11696581","id":"PMC_11696581","title":"The forkhead transcription factor Foxo1 (Fkhr) confers insulin sensitivity onto glucose-6-phosphatase expression.","date":"2001","source":"The Journal of clinical investigation","url":"https://pubmed.ncbi.nlm.nih.gov/11696581","citation_count":495,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"16154098","id":"PMC_16154098","title":"FoxO1 protects against pancreatic beta cell failure through NeuroD and MafA induction.","date":"2005","source":"Cell metabolism","url":"https://pubmed.ncbi.nlm.nih.gov/16154098","citation_count":492,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"13679577","id":"PMC_13679577","title":"Insulin-induced phosphorylation of FKHR (Foxo1) targets to proteasomal degradation.","date":"2003","source":"Proceedings of the National Academy of Sciences of the United States of America","url":"https://pubmed.ncbi.nlm.nih.gov/13679577","citation_count":439,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"16076959","id":"PMC_16076959","title":"Acetylation of Foxo1 alters its DNA-binding ability and sensitivity to phosphorylation.","date":"2005","source":"Proceedings of the National Academy of Sciences of the United States of America","url":"https://pubmed.ncbi.nlm.nih.gov/16076959","citation_count":419,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"12150827","id":"PMC_12150827","title":"A novel mechanism of gene regulation and tumor suppression by the transcription factor FKHR.","date":"2002","source":"Cancer cell","url":"https://pubmed.ncbi.nlm.nih.gov/12150827","citation_count":363,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"29158051","id":"PMC_29158051","title":"The regulation of FOXO1 and its role in disease progression.","date":"2017","source":"Life sciences","url":"https://pubmed.ncbi.nlm.nih.gov/29158051","citation_count":340,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"9479491","id":"PMC_9479491","title":"Cloning and characterization of three human forkhead genes that comprise an FKHR-like gene subfamily.","date":"1998","source":"Genomics","url":"https://pubmed.ncbi.nlm.nih.gov/9479491","citation_count":297,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"15184386","id":"PMC_15184386","title":"Abnormal angiogenesis in Foxo1 (Fkhr)-deficient mice.","date":"2004","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/15184386","citation_count":284,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"28446439","id":"PMC_28446439","title":"PAX3-FOXO1 Establishes Myogenic Super Enhancers and Confers BET Bromodomain Vulnerability.","date":"2017","source":"Cancer discovery","url":"https://pubmed.ncbi.nlm.nih.gov/28446439","citation_count":258,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"14710188","id":"PMC_14710188","title":"D4476, a cell-permeant inhibitor of CK1, suppresses the site-specific phosphorylation and nuclear exclusion of FOXO1a.","date":"2004","source":"EMBO reports","url":"https://pubmed.ncbi.nlm.nih.gov/14710188","citation_count":227,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"32802173","id":"PMC_32802173","title":"METTL14 aggravates endothelial inflammation and atherosclerosis by increasing FOXO1 N6-methyladeosine modifications.","date":"2020","source":"Theranostics","url":"https://pubmed.ncbi.nlm.nih.gov/32802173","citation_count":224,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"11311120","id":"PMC_11311120","title":"The kinase DYRK1A phosphorylates the transcription factor FKHR at Ser329 in vitro, a novel in vivo phosphorylation site.","date":"2001","source":"The Biochemical journal","url":"https://pubmed.ncbi.nlm.nih.gov/11311120","citation_count":218,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"21045807","id":"PMC_21045807","title":"FoxO1 regulates Tlr4 inflammatory pathway signalling in macrophages.","date":"2010","source":"The EMBO journal","url":"https://pubmed.ncbi.nlm.nih.gov/21045807","citation_count":212,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"15520281","id":"PMC_15520281","title":"Pax3:Fkhr interferes with embryonic Pax3 and Pax7 function: implications for alveolar rhabdomyosarcoma cell of origin.","date":"2004","source":"Genes & development","url":"https://pubmed.ncbi.nlm.nih.gov/15520281","citation_count":196,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"10393185","id":"PMC_10393185","title":"The Pax3-FKHR oncoprotein is unresponsive to the Pax3-associated repressor hDaxx.","date":"1999","source":"The EMBO journal","url":"https://pubmed.ncbi.nlm.nih.gov/10393185","citation_count":183,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"22147007","id":"PMC_22147007","title":"Hepatic suppression of Foxo1 and Foxo3 causes hypoglycemia and hyperlipidemia in mice.","date":"2011","source":"Endocrinology","url":"https://pubmed.ncbi.nlm.nih.gov/22147007","citation_count":146,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"12482965","id":"PMC_12482965","title":"AKT-independent protection of prostate cancer cells from apoptosis mediated through complex formation between the androgen receptor and FKHR.","date":"2003","source":"Molecular and cellular biology","url":"https://pubmed.ncbi.nlm.nih.gov/12482965","citation_count":137,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"18457914","id":"PMC_18457914","title":"PAX3-FOXO1 fusion gene in rhabdomyosarcoma.","date":"2008","source":"Cancer letters","url":"https://pubmed.ncbi.nlm.nih.gov/18457914","citation_count":136,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"8789435","id":"PMC_8789435","title":"In vivo amplification of the PAX3-FKHR and PAX7-FKHR fusion genes in alveolar rhabdomyosarcoma.","date":"1996","source":"Human molecular genetics","url":"https://pubmed.ncbi.nlm.nih.gov/8789435","citation_count":132,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"33539225","id":"PMC_33539225","title":"KLF5 Is Induced by FOXO1 and Causes Oxidative Stress and Diabetic Cardiomyopathy.","date":"2020","source":"Circulation research","url":"https://pubmed.ncbi.nlm.nih.gov/33539225","citation_count":122,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"23263330","id":"PMC_23263330","title":"FoxO1 is crucial for sustaining cardiomyocyte metabolism and cell survival.","date":"2012","source":"Cardiovascular research","url":"https://pubmed.ncbi.nlm.nih.gov/23263330","citation_count":119,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"16849544","id":"PMC_16849544","title":"FOXO1A is a candidate for the 13q14 tumor suppressor gene inhibiting androgen receptor signaling in prostate cancer.","date":"2006","source":"Cancer research","url":"https://pubmed.ncbi.nlm.nih.gov/16849544","citation_count":116,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"31849924","id":"PMC_31849924","title":"Mucosal Immunity and the FOXO1 Transcription Factors.","date":"2019","source":"Frontiers in immunology","url":"https://pubmed.ncbi.nlm.nih.gov/31849924","citation_count":108,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"18972406","id":"PMC_18972406","title":"Selenoprotein P expression is controlled through interaction of the coactivator PGC-1alpha with FoxO1a and hepatocyte nuclear factor 4alpha transcription factors.","date":"2008","source":"Hepatology (Baltimore, Md.)","url":"https://pubmed.ncbi.nlm.nih.gov/18972406","citation_count":104,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"8790412","id":"PMC_8790412","title":"The hybrid PAX3-FKHR fusion protein of alveolar rhabdomyosarcoma transforms fibroblasts in culture.","date":"1996","source":"Proceedings of the National Academy of Sciences of the United States of America","url":"https://pubmed.ncbi.nlm.nih.gov/8790412","citation_count":103,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"25378413","id":"PMC_25378413","title":"Inhibition of FOXO1/3 promotes vascular calcification.","date":"2014","source":"Arteriosclerosis, thrombosis, and vascular biology","url":"https://pubmed.ncbi.nlm.nih.gov/25378413","citation_count":102,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"32057903","id":"PMC_32057903","title":"Resveratrol promotes osteogenesis via activating SIRT1/FoxO1 pathway in osteoporosis mice.","date":"2020","source":"Life sciences","url":"https://pubmed.ncbi.nlm.nih.gov/32057903","citation_count":91,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"25226535","id":"PMC_25226535","title":"FOXO1, TGF-β regulation and wound healing.","date":"2014","source":"International journal of molecular sciences","url":"https://pubmed.ncbi.nlm.nih.gov/25226535","citation_count":89,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"17993506","id":"PMC_17993506","title":"FOXC1 is required for cell viability and resistance to oxidative stress in the eye through the transcriptional regulation of FOXO1A.","date":"2007","source":"Human molecular genetics","url":"https://pubmed.ncbi.nlm.nih.gov/17993506","citation_count":89,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"17245409","id":"PMC_17245409","title":"Haplotypes in the human Foxo1a and Foxo3a genes; impact on disease and mortality at old age.","date":"2007","source":"European journal of human genetics : EJHG","url":"https://pubmed.ncbi.nlm.nih.gov/17245409","citation_count":89,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"12560069","id":"PMC_12560069","title":"Estrogen regulation of Pak1 and FKHR pathways in breast cancer cells.","date":"2003","source":"FEBS letters","url":"https://pubmed.ncbi.nlm.nih.gov/12560069","citation_count":87,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"18022385","id":"PMC_18022385","title":"Effects of PAX3-FKHR on malignant phenotypes in alveolar rhabdomyosarcoma.","date":"2007","source":"Biochemical and biophysical research communications","url":"https://pubmed.ncbi.nlm.nih.gov/18022385","citation_count":85,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"19727442","id":"PMC_19727442","title":"HoxA-11 and FOXO1A cooperate to regulate decidual prolactin expression: towards inferring the core transcriptional regulators of decidual genes.","date":"2009","source":"PloS one","url":"https://pubmed.ncbi.nlm.nih.gov/19727442","citation_count":79,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"17030088","id":"PMC_17030088","title":"APPL1, APPL2, Akt2 and FOXO1a interact with FSHR in a potential signaling complex.","date":"2006","source":"Molecular and cellular endocrinology","url":"https://pubmed.ncbi.nlm.nih.gov/17030088","citation_count":78,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"30780132","id":"PMC_30780132","title":"Molecular mechanisms of FOXO1 in adipocyte differentiation.","date":"2019","source":"Journal of molecular endocrinology","url":"https://pubmed.ncbi.nlm.nih.gov/30780132","citation_count":76,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"26470727","id":"PMC_26470727","title":"Functional regulation of FoxO1 in neural stem cell differentiation.","date":"2015","source":"Cell death and differentiation","url":"https://pubmed.ncbi.nlm.nih.gov/26470727","citation_count":76,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"29574902","id":"PMC_29574902","title":"FOXO1 regulates VEGFA expression and promotes angiogenesis in healing wounds.","date":"2018","source":"The Journal of pathology","url":"https://pubmed.ncbi.nlm.nih.gov/29574902","citation_count":74,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"33075563","id":"PMC_33075563","title":"S100A11 Promotes Liver Steatosis via FOXO1-Mediated Autophagy and Lipogenesis.","date":"2020","source":"Cellular and molecular gastroenterology and hepatology","url":"https://pubmed.ncbi.nlm.nih.gov/33075563","citation_count":72,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"33503413","id":"PMC_33503413","title":"FOXO1 constrains activation and regulates senescence in CD8 T cells.","date":"2021","source":"Cell reports","url":"https://pubmed.ncbi.nlm.nih.gov/33503413","citation_count":69,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"7630639","id":"PMC_7630639","title":"The alveolar rhabdomyosarcoma PAX3/FKHR fusion protein is a transcriptional activator.","date":"1995","source":"Oncogene","url":"https://pubmed.ncbi.nlm.nih.gov/7630639","citation_count":67,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"20151947","id":"PMC_20151947","title":"FoxO1 - the key for the pathogenesis and therapy of acne?","date":"2010","source":"Journal der Deutschen Dermatologischen Gesellschaft = Journal of the German Society of Dermatology : JDDG","url":"https://pubmed.ncbi.nlm.nih.gov/20151947","citation_count":66,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"16614392","id":"PMC_16614392","title":"Forkhead transcription factor FOXO1A is critical for induction of human decidualization.","date":"2006","source":"The Journal of endocrinology","url":"https://pubmed.ncbi.nlm.nih.gov/16614392","citation_count":66,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"25918228","id":"PMC_25918228","title":"FOXO1 differentially regulates both normal and diabetic wound healing.","date":"2015","source":"The Journal of cell biology","url":"https://pubmed.ncbi.nlm.nih.gov/25918228","citation_count":66,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"11549601","id":"PMC_11549601","title":"PAX3-FKHR induces morphological change and enhances cellular proliferation and invasion in rhabdomyosarcoma.","date":"2001","source":"The American journal of pathology","url":"https://pubmed.ncbi.nlm.nih.gov/11549601","citation_count":64,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"18202777","id":"PMC_18202777","title":"Overexpression of phospho-Akt correlates with phosphorylation of EGF receptor, FKHR and BAD in nasopharyngeal carcinoma.","date":"2008","source":"Oncology reports","url":"https://pubmed.ncbi.nlm.nih.gov/18202777","citation_count":64,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"21196578","id":"PMC_21196578","title":"FoxO1 and SIRT1 regulate beta-cell responses to nitric oxide.","date":"2011","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/21196578","citation_count":62,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"22438832","id":"PMC_22438832","title":"The yeast forkhead transcription factors fkh1 and fkh2 regulate lifespan and stress response together with the anaphase-promoting complex.","date":"2012","source":"PLoS genetics","url":"https://pubmed.ncbi.nlm.nih.gov/22438832","citation_count":59,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"30552714","id":"PMC_30552714","title":"PAR2 promotes M1 macrophage polarization and inflammation via FOXO1 pathway.","date":"2018","source":"Journal of cellular biochemistry","url":"https://pubmed.ncbi.nlm.nih.gov/30552714","citation_count":59,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"22710712","id":"PMC_22710712","title":"Alveolar rhabdomyosarcoma-associated proteins PAX3/FOXO1A and PAX7/FOXO1A suppress the transcriptional activity of MyoD-target genes in muscle stem cells.","date":"2012","source":"Oncogene","url":"https://pubmed.ncbi.nlm.nih.gov/22710712","citation_count":58,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"11863357","id":"PMC_11863357","title":"Identification of target genes regulated by PAX3 and PAX3-FKHR in embryogenesis and alveolar rhabdomyosarcoma.","date":"2002","source":"Genomics","url":"https://pubmed.ncbi.nlm.nih.gov/11863357","citation_count":57,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"32244542","id":"PMC_32244542","title":"Adipose Tissue and FoxO1: Bridging Physiology and Mechanisms.","date":"2020","source":"Cells","url":"https://pubmed.ncbi.nlm.nih.gov/32244542","citation_count":56,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"35342363","id":"PMC_35342363","title":"HB-EGF induces mitochondrial dysfunction via estrogen hypersecretion in granulosa cells dependent on cAMP-PKA-JNK/ERK-Ca2+-FOXO1 pathway.","date":"2022","source":"International journal of biological sciences","url":"https://pubmed.ncbi.nlm.nih.gov/35342363","citation_count":56,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"34287641","id":"PMC_34287641","title":"Foxo1 controls gut homeostasis and commensalism by regulating mucus secretion.","date":"2021","source":"The Journal of experimental medicine","url":"https://pubmed.ncbi.nlm.nih.gov/34287641","citation_count":53,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"29988091","id":"PMC_29988091","title":"Maintenance of CD4 T cell fitness through regulation of Foxo1.","date":"2018","source":"Nature immunology","url":"https://pubmed.ncbi.nlm.nih.gov/29988091","citation_count":53,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"35700043","id":"PMC_35700043","title":"Myeloid FoxO1 depletion attenuates hepatic inflammation and prevents nonalcoholic steatohepatitis.","date":"2022","source":"The Journal of clinical investigation","url":"https://pubmed.ncbi.nlm.nih.gov/35700043","citation_count":52,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"22496671","id":"PMC_22496671","title":"Regulation of budding yeast mating-type switching donor preference by the FHA domain of Fkh1.","date":"2012","source":"PLoS genetics","url":"https://pubmed.ncbi.nlm.nih.gov/22496671","citation_count":52,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"17638879","id":"PMC_17638879","title":"The PAX3-FKHR fusion gene of rhabdomyosarcoma cooperates with loss of p16INK4A to promote bypass of cellular senescence.","date":"2007","source":"Cancer research","url":"https://pubmed.ncbi.nlm.nih.gov/17638879","citation_count":52,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"8634710","id":"PMC_8634710","title":"Structural characterization of the FKHR gene and its rearrangement in alveolar rhabdomyosarcoma.","date":"1995","source":"Human molecular genetics","url":"https://pubmed.ncbi.nlm.nih.gov/8634710","citation_count":51,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"17024043","id":"PMC_17024043","title":"Gluconeogenesis: re-evaluating the FOXO1-PGC-1alpha connection.","date":"2006","source":"Nature","url":"https://pubmed.ncbi.nlm.nih.gov/17024043","citation_count":51,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"25786691","id":"PMC_25786691","title":"FOXO1 regulates dendritic cell activity through ICAM-1 and CCR7.","date":"2015","source":"Journal of immunology (Baltimore, Md. : 1950)","url":"https://pubmed.ncbi.nlm.nih.gov/25786691","citation_count":50,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"28499891","id":"PMC_28499891","title":"SAPHO, autophagy, IL-1, FoxO1, and Propionibacterium (Cutibacterium) acnes.","date":"2017","source":"Joint bone spine","url":"https://pubmed.ncbi.nlm.nih.gov/28499891","citation_count":48,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"27260854","id":"PMC_27260854","title":"FoxO1 antagonist suppresses autophagy and lipid droplet growth in adipocytes.","date":"2016","source":"Cell cycle (Georgetown, Tex.)","url":"https://pubmed.ncbi.nlm.nih.gov/27260854","citation_count":47,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"8889501","id":"PMC_8889501","title":"Novel formation and amplification of the PAX7-FKHR fusion gene in a case of alveolar rhabdomyosarcoma.","date":"1996","source":"Genes, chromosomes & cancer","url":"https://pubmed.ncbi.nlm.nih.gov/8889501","citation_count":46,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"20061393","id":"PMC_20061393","title":"Regulation of FOXO1 by TAK1-Nemo-like kinase pathway.","date":"2010","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/20061393","citation_count":45,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"25157276","id":"PMC_25157276","title":"Down Regulation of FOXO1 Promotes Cell Proliferation in Cervical Cancer.","date":"2014","source":"Journal of Cancer","url":"https://pubmed.ncbi.nlm.nih.gov/25157276","citation_count":44,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"20449867","id":"PMC_20449867","title":"Foxo1 regulates marginal zone B-cell development.","date":"2010","source":"European journal of immunology","url":"https://pubmed.ncbi.nlm.nih.gov/20449867","citation_count":42,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"12196101","id":"PMC_12196101","title":"Effect of multiple phosphorylation events on the transcription factors FKHR, FKHRL1 and AFX.","date":"2002","source":"Biochemical Society transactions","url":"https://pubmed.ncbi.nlm.nih.gov/12196101","citation_count":41,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"16287709","id":"PMC_16287709","title":"Coordinated oncogenic transformation and inhibition of host immune responses by the PAX3-FKHR fusion oncoprotein.","date":"2005","source":"The Journal of experimental medicine","url":"https://pubmed.ncbi.nlm.nih.gov/16287709","citation_count":41,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"12183363","id":"PMC_12183363","title":"Saccharomyces forkhead protein Fkh1 regulates donor preference during mating-type switching through the recombination enhancer.","date":"2002","source":"Genes & development","url":"https://pubmed.ncbi.nlm.nih.gov/12183363","citation_count":40,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"26728715","id":"PMC_26728715","title":"Quantitative BrdU immunoprecipitation method demonstrates that Fkh1 and Fkh2 are rate-limiting activators of replication origins that reprogram replication timing in G1 phase.","date":"2016","source":"Genome research","url":"https://pubmed.ncbi.nlm.nih.gov/26728715","citation_count":38,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"22533991","id":"PMC_22533991","title":"Carnitine palmitoyltransferase 1A (CPT1A): a transcriptional target of PAX3-FKHR and mediates PAX3-FKHR-dependent motility in alveolar rhabdomyosarcoma cells.","date":"2012","source":"BMC cancer","url":"https://pubmed.ncbi.nlm.nih.gov/22533991","citation_count":37,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"33089967","id":"PMC_33089967","title":"The role and molecular mechanism of FoxO1 in mediating cardiac hypertrophy.","date":"2020","source":"ESC heart failure","url":"https://pubmed.ncbi.nlm.nih.gov/33089967","citation_count":35,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"35857643","id":"PMC_35857643","title":"Targeting KDM4 for treating PAX3-FOXO1-driven alveolar rhabdomyosarcoma.","date":"2022","source":"Science translational medicine","url":"https://pubmed.ncbi.nlm.nih.gov/35857643","citation_count":34,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"26962001","id":"PMC_26962001","title":"HDAC2 regulates FoxO1 during RANKL-induced osteoclastogenesis.","date":"2016","source":"American journal of physiology. Cell physiology","url":"https://pubmed.ncbi.nlm.nih.gov/26962001","citation_count":34,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"16020479","id":"PMC_16020479","title":"Regulation of the proapoptotic factor FOXO1 (FKHR) in cardiomyocytes by growth factors and alpha1-adrenergic agonists.","date":"2005","source":"Endocrinology","url":"https://pubmed.ncbi.nlm.nih.gov/16020479","citation_count":34,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"27329843","id":"PMC_27329843","title":"Aquaporin 9 inhibits hepatocellular carcinoma through up-regulating FOXO1 expression.","date":"2016","source":"Oncotarget","url":"https://pubmed.ncbi.nlm.nih.gov/27329843","citation_count":33,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"27864345","id":"PMC_27864345","title":"PAX3-FOXO1A Expression in Rhabdomyosarcoma Is Driven by the Targetable Nuclear Receptor NR4A1.","date":"2016","source":"Cancer research","url":"https://pubmed.ncbi.nlm.nih.gov/27864345","citation_count":31,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"25694609","id":"PMC_25694609","title":"FOXO1 mediates RANKL-induced osteoclast formation and activity.","date":"2015","source":"Journal of immunology (Baltimore, Md. : 1950)","url":"https://pubmed.ncbi.nlm.nih.gov/25694609","citation_count":31,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"29339772","id":"PMC_29339772","title":"Foxo1 Promotes Th9 Cell Differentiation and Airway Allergy.","date":"2018","source":"Scientific reports","url":"https://pubmed.ncbi.nlm.nih.gov/29339772","citation_count":30,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"22710718","id":"PMC_22710718","title":"P-cadherin is a direct PAX3-FOXO1A target involved in alveolar rhabdomyosarcoma aggressiveness.","date":"2012","source":"Oncogene","url":"https://pubmed.ncbi.nlm.nih.gov/22710718","citation_count":30,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"37506793","id":"PMC_37506793","title":"FOXO1 regulates the formation of bovine fat by targeting CD36 and STEAP4.","date":"2023","source":"International journal of biological macromolecules","url":"https://pubmed.ncbi.nlm.nih.gov/37506793","citation_count":30,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"17350164","id":"PMC_17350164","title":"PAX3 and PAX3-FKHR promote rhabdomyosarcoma cell survival through downregulation of PTEN.","date":"2007","source":"Cancer letters","url":"https://pubmed.ncbi.nlm.nih.gov/17350164","citation_count":29,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"30697272","id":"PMC_30697272","title":"Trichostatin A activates FOXO1 and induces autophagy in osteosarcoma.","date":"2018","source":"Archives of medical science : AMS","url":"https://pubmed.ncbi.nlm.nih.gov/30697272","citation_count":29,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"33493902","id":"PMC_33493902","title":"Galanin promotes autophagy and alleviates apoptosis in the hypertrophied heart through FoxO1 pathway.","date":"2021","source":"Redox biology","url":"https://pubmed.ncbi.nlm.nih.gov/33493902","citation_count":29,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"31026811","id":"PMC_31026811","title":"Glucagon regulates hepatic mitochondrial function and biogenesis through FOXO1.","date":"2019","source":"The Journal of endocrinology","url":"https://pubmed.ncbi.nlm.nih.gov/31026811","citation_count":29,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"34385359","id":"PMC_34385359","title":"Akt Regulates Sox10 Expression to Control Oligodendrocyte Differentiation via Phosphorylating FoxO1.","date":"2021","source":"The Journal of neuroscience : the official journal of the Society for Neuroscience","url":"https://pubmed.ncbi.nlm.nih.gov/34385359","citation_count":28,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"34419603","id":"PMC_34419603","title":"TGFβ1 signaling protects chondrocytes against oxidative stress via FOXO1-autophagy axis.","date":"2021","source":"Osteoarthritis and cartilage","url":"https://pubmed.ncbi.nlm.nih.gov/34419603","citation_count":28,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"37602516","id":"PMC_37602516","title":"Suppression of FOXO1 attenuates inflamm-aging and improves liver function during aging.","date":"2023","source":"Aging cell","url":"https://pubmed.ncbi.nlm.nih.gov/37602516","citation_count":27,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"28790135","id":"PMC_28790135","title":"DDB1-Mediated CRY1 Degradation Promotes FOXO1-Driven Gluconeogenesis in Liver.","date":"2017","source":"Diabetes","url":"https://pubmed.ncbi.nlm.nih.gov/28790135","citation_count":27,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"16107711","id":"PMC_16107711","title":"FoxO1a-cyclic GMP-dependent kinase I interactions orchestrate myoblast fusion.","date":"2005","source":"Molecular and cellular biology","url":"https://pubmed.ncbi.nlm.nih.gov/16107711","citation_count":27,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"30144329","id":"PMC_30144329","title":"FoxO1 enhances differentiation and apoptosis in human primary keratinocytes.","date":"2018","source":"Experimental dermatology","url":"https://pubmed.ncbi.nlm.nih.gov/30144329","citation_count":26,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"19837876","id":"PMC_19837876","title":"Atrial natriuretic peptide promotes pancreatic islet beta-cell growth and Akt/Foxo1a/cyclin D2 signaling.","date":"2009","source":"Endocrinology","url":"https://pubmed.ncbi.nlm.nih.gov/19837876","citation_count":25,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"33627785","id":"PMC_33627785","title":"FOXF1 is required for the oncogenic properties of PAX3-FOXO1 in rhabdomyosarcoma.","date":"2021","source":"Oncogene","url":"https://pubmed.ncbi.nlm.nih.gov/33627785","citation_count":25,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"18293098","id":"PMC_18293098","title":"Tissue expression of porcine FoxO1 and its negative regulation during primary preadipocyte differentiation.","date":"2008","source":"Molecular biology reports","url":"https://pubmed.ncbi.nlm.nih.gov/18293098","citation_count":25,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"28851713","id":"PMC_28851713","title":"SCP4 Promotes Gluconeogenesis Through FoxO1/3a Dephosphorylation.","date":"2017","source":"Diabetes","url":"https://pubmed.ncbi.nlm.nih.gov/28851713","citation_count":24,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"19150739","id":"PMC_19150739","title":"Localisation and expression of FoxO1 proteins in human gestational tissues.","date":"2009","source":"Placenta","url":"https://pubmed.ncbi.nlm.nih.gov/19150739","citation_count":23,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"17457058","id":"PMC_17457058","title":"CDK2 and FOXO1: a fork in the road for cell fate decisions.","date":"2007","source":"Cell cycle (Georgetown, Tex.)","url":"https://pubmed.ncbi.nlm.nih.gov/17457058","citation_count":23,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"22333587","id":"PMC_22333587","title":"AKT and PAX3-FKHR cooperation enforces myogenic differentiation blockade in alveolar rhabdomyosarcoma cell.","date":"2012","source":"Cell cycle (Georgetown, Tex.)","url":"https://pubmed.ncbi.nlm.nih.gov/22333587","citation_count":22,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"15109499","id":"PMC_15109499","title":"Foxo transcription factors induce the atrophy-related ubiquitin ligase atrogin-1 and cause skeletal muscle atrophy.","date":"2004","source":"Cell","url":"https://pubmed.ncbi.nlm.nih.gov/15109499","citation_count":2402,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"12477932","id":"PMC_12477932","title":"Generation and initial analysis of more than 15,000 full-length human and mouse cDNA sequences.","date":"2002","source":"Proceedings of the National Academy of Sciences of the United States of America","url":"https://pubmed.ncbi.nlm.nih.gov/12477932","citation_count":1479,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"10783894","id":"PMC_10783894","title":"AFX-like Forkhead transcription factors mediate cell-cycle regulation by Ras and PKB through p27kip1.","date":"2000","source":"Nature","url":"https://pubmed.ncbi.nlm.nih.gov/10783894","citation_count":1227,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"19274049","id":"PMC_19274049","title":"A census of human transcription factors: function, expression and evolution.","date":"2009","source":"Nature reviews. Genetics","url":"https://pubmed.ncbi.nlm.nih.gov/19274049","citation_count":1191,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"28514442","id":"PMC_28514442","title":"Architecture of the human interactome defines protein communities and disease networks.","date":"2017","source":"Nature","url":"https://pubmed.ncbi.nlm.nih.gov/28514442","citation_count":1085,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"21440011","id":"PMC_21440011","title":"Akt, FoxO and regulation of apoptosis.","date":"2011","source":"Biochimica et biophysica acta","url":"https://pubmed.ncbi.nlm.nih.gov/21440011","citation_count":922,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"15084259","id":"PMC_15084259","title":"Integration of Smad and forkhead pathways in the control of neuroepithelial and glioblastoma cell proliferation.","date":"2004","source":"Cell","url":"https://pubmed.ncbi.nlm.nih.gov/15084259","citation_count":837,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"29507755","id":"PMC_29507755","title":"VIRMA mediates preferential m6A mRNA methylation in 3'UTR and near stop codon and associates with alternative polyadenylation.","date":"2018","source":"Cell discovery","url":"https://pubmed.ncbi.nlm.nih.gov/29507755","citation_count":829,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"8275086","id":"PMC_8275086","title":"Fusion of a fork head domain gene to PAX3 in the solid tumour alveolar rhabdomyosarcoma.","date":"1993","source":"Nature genetics","url":"https://pubmed.ncbi.nlm.nih.gov/8275086","citation_count":815,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"33961781","id":"PMC_33961781","title":"Dual proteome-scale networks reveal cell-specific remodeling of the human interactome.","date":"2021","source":"Cell","url":"https://pubmed.ncbi.nlm.nih.gov/33961781","citation_count":705,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"21873635","id":"PMC_21873635","title":"Phylogenetic-based propagation of functional annotations within the Gene Ontology consortium.","date":"2011","source":"Briefings in bioinformatics","url":"https://pubmed.ncbi.nlm.nih.gov/21873635","citation_count":656,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"15905404","id":"PMC_15905404","title":"Functional interaction between beta-catenin and FOXO in oxidative stress signaling.","date":"2005","source":"Science (New York, N.Y.)","url":"https://pubmed.ncbi.nlm.nih.gov/15905404","citation_count":621,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"21708191","id":"PMC_21708191","title":"FoxO transcription factors; Regulation by AKT and 14-3-3 proteins.","date":"2011","source":"Biochimica et biophysica acta","url":"https://pubmed.ncbi.nlm.nih.gov/21708191","citation_count":593,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"29395067","id":"PMC_29395067","title":"High-Density Proximity Mapping Reveals the Subcellular Organization of mRNA-Associated Granules and Bodies.","date":"2018","source":"Molecular cell","url":"https://pubmed.ncbi.nlm.nih.gov/29395067","citation_count":580,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"12039929","id":"PMC_12039929","title":"PAX3-FKHR and PAX7-FKHR gene fusions are prognostic indicators in alveolar rhabdomyosarcoma: a report from the children's oncology group.","date":"2002","source":"Journal of clinical oncology : official journal of the American Society of Clinical Oncology","url":"https://pubmed.ncbi.nlm.nih.gov/12039929","citation_count":558,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"20543840","id":"PMC_20543840","title":"Cytosolic FoxO1 is essential for the induction of autophagy and tumour suppressor activity.","date":"2010","source":"Nature cell biology","url":"https://pubmed.ncbi.nlm.nih.gov/20543840","citation_count":519,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"15220471","id":"PMC_15220471","title":"Silent information regulator 2 potentiates Foxo1-mediated transcription through its deacetylase activity.","date":"2004","source":"Proceedings of the National Academy of Sciences of the United States of America","url":"https://pubmed.ncbi.nlm.nih.gov/15220471","citation_count":504,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"19574223","id":"PMC_19574223","title":"Coordinate regulation of FOXO1 by miR-27a, miR-96, and miR-182 in breast cancer cells.","date":"2009","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/19574223","citation_count":497,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"26735015","id":"PMC_26735015","title":"FOXO1 couples metabolic activity and growth state in the vascular endothelium.","date":"2016","source":"Nature","url":"https://pubmed.ncbi.nlm.nih.gov/26735015","citation_count":479,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"16100571","id":"PMC_16100571","title":"Involvement of Foxo transcription factors in angiogenesis and postnatal neovascularization.","date":"2005","source":"The Journal of clinical investigation","url":"https://pubmed.ncbi.nlm.nih.gov/16100571","citation_count":456,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"18235501","id":"PMC_18235501","title":"DBC1 is a negative regulator of SIRT1.","date":"2008","source":"Nature","url":"https://pubmed.ncbi.nlm.nih.gov/18235501","citation_count":445,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"15668399","id":"PMC_15668399","title":"Skp2 inhibits FOXO1 in tumor suppression through ubiquitin-mediated degradation.","date":"2005","source":"Proceedings of the National Academy of Sciences of the United States of America","url":"https://pubmed.ncbi.nlm.nih.gov/15668399","citation_count":444,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"15489334","id":"PMC_15489334","title":"The status, quality, and expansion of the NIH full-length cDNA project: the Mammalian Gene Collection (MGC).","date":"2004","source":"Genome research","url":"https://pubmed.ncbi.nlm.nih.gov/15489334","citation_count":438,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"16492665","id":"PMC_16492665","title":"FoxO1 regulates multiple metabolic pathways in the liver: effects on gluconeogenic, glycolytic, and lipogenic gene expression.","date":"2006","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/16492665","citation_count":433,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"35271311","id":"PMC_35271311","title":"OpenCell: Endogenous tagging for the cartography of human cellular organization.","date":"2022","source":"Science (New York, N.Y.)","url":"https://pubmed.ncbi.nlm.nih.gov/35271311","citation_count":432,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"18951090","id":"PMC_18951090","title":"Arginine methylation of FOXO transcription factors inhibits their phosphorylation by Akt.","date":"2008","source":"Molecular cell","url":"https://pubmed.ncbi.nlm.nih.gov/18951090","citation_count":378,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"20351326","id":"PMC_20351326","title":"Fusion gene-negative alveolar rhabdomyosarcoma is clinically and molecularly indistinguishable from embryonal rhabdomyosarcoma.","date":"2010","source":"Journal of clinical oncology : official journal of the American Society of Clinical Oncology","url":"https://pubmed.ncbi.nlm.nih.gov/20351326","citation_count":377,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"16753578","id":"PMC_16753578","title":"GCN5 acetyltransferase complex controls glucose metabolism through transcriptional repression of PGC-1alpha.","date":"2006","source":"Cell metabolism","url":"https://pubmed.ncbi.nlm.nih.gov/16753578","citation_count":372,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"24140020","id":"PMC_24140020","title":"mTOR complex 2 controls glycolytic metabolism in glioblastoma through FoxO acetylation and upregulation of c-Myc.","date":"2013","source":"Cell metabolism","url":"https://pubmed.ncbi.nlm.nih.gov/24140020","citation_count":363,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"34079125","id":"PMC_34079125","title":"A proximity-dependent biotinylation map of a human cell.","date":"2021","source":"Nature","url":"https://pubmed.ncbi.nlm.nih.gov/34079125","citation_count":339,"is_preprint":false,"source_track":"gene2pubmed"}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":48418,"output_tokens":6660,"usd":0.122577},"stage2":{"model":"claude-opus-4-6","input_tokens":10357,"output_tokens":4101,"usd":0.231465},"total_usd":0.830922,"stage1_batch_id":"msgbatch_0118RScg3WGaGkaZRtWZbtce","stage2_batch_id":"msgbatch_01Cpfzvtbj42aqp5oYZ5Te2s","note":"batch pricing = 50% of standard","round2_stage1":{"model":"claude-sonnet-4-6","input_tokens":61105,"output_tokens":10814,"usd":0.172763},"round2_rules_fired":"R3","round2_stage2":{"model":"claude-opus-4-6","input_tokens":15014,"output_tokens":5107,"usd":0.304117}},"stage1_raw":"```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 1999,\n      \"finding\": \"Akt (PKB) directly phosphorylates FOXO1 (FKHR) at three sites (Thr-24, Ser-256, Ser-319) in vitro and in vivo, suppressing FOXO1-mediated transcriptional activation and promoting cytoplasmic retention; mutation of all three sites to alanine renders FOXO1 resistant to Akt inhibition and causes apoptosis in a DNA-binding-dependent manner.\",\n      \"method\": \"In vitro kinase assay, in vivo phosphorylation in 293T cells, site-directed mutagenesis, transcriptional reporter assay\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 — in vitro reconstitution and mutagenesis, independently replicated in two papers (PMID:10358014 and PMID:10358075)\",\n      \"pmids\": [\"10358014\", \"10358075\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 1999,\n      \"finding\": \"PKBα phosphorylates FOXO1 at Thr-24, Ser-256, and Ser-319, which all lie within the canonical PKB consensus (Arg-Xaa-Arg-Xaa-Xaa-Ser/Thr); IGF-1-induced phosphorylation of these sites is abolished by PI3K inhibitor wortmannin but not by MEK or mTOR inhibitors, placing FOXO1 downstream of the PI3K-PDK1-PKB cascade.\",\n      \"method\": \"In vitro kinase assay, co-transfection with PKB/PDK1, pharmacological inhibition in 293 cells\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 — in vitro assay plus pharmacological pathway epistasis, replicated\",\n      \"pmids\": [\"10358075\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2001,\n      \"finding\": \"FOXO1 binds to the insulin-responsive sequence in the glucose-6-phosphatase (G6P) promoter and is required for insulin to suppress glucocorticoid/cAMP-induced G6P expression in liver/kidney cells; dominant-negative FOXO1 partially inhibits G6P and PEPCK expression, establishing FOXO1 as the transcription factor mediating insulin regulation of hepatic gluconeogenesis.\",\n      \"method\": \"Recombinant adenovirus transduction, dominant-negative mutant expression, reporter assay in primary hepatocytes and kidney cells\",\n      \"journal\": \"The Journal of clinical investigation\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — clean gain- and loss-of-function with defined transcriptional readout\",\n      \"pmids\": [\"11696581\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2001,\n      \"finding\": \"DYRK1A phosphorylates FOXO1 at Ser-329 in vitro and co-localizes and co-immunoprecipitates with FOXO1 in cell extracts; Ser-329 phosphorylation reduces nuclear FOXO1 abundance and attenuates transcriptional activation independently of the PI3K/PKB pathway.\",\n      \"method\": \"Biochemical purification of kinase from rabbit skeletal muscle, in vitro kinase assay, co-immunoprecipitation, co-localization imaging\",\n      \"journal\": \"The Biochemical journal\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 — purification to near-homogeneity plus in vitro assay and co-IP\",\n      \"pmids\": [\"11311120\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2002,\n      \"finding\": \"FOXO1 suppresses cell cycle progression by transcriptionally repressing D-type cyclins rather than by activating IRS-regulated genes; chromatin immunoprecipitation confirms FOXO1 occupancy at cyclin D loci, and this repressive mechanism is required for FOXO1-mediated inhibition of transformation.\",\n      \"method\": \"Transcriptional profiling, chromatin immunoprecipitation, functional transformation assays\",\n      \"journal\": \"Cancer cell\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — ChIP plus functional epistasis, multiple orthogonal methods\",\n      \"pmids\": [\"12150827\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2003,\n      \"finding\": \"Insulin-induced, PI3K/PKB-dependent phosphorylation of FOXO1 triggers its ubiquitination and proteasomal degradation; efficient ubiquitination requires both phosphorylation at Thr-24/Ser-256/Ser-319 and cytoplasmic retention of FOXO1.\",\n      \"method\": \"Proteasome inhibitor treatment, in vivo and in vitro ubiquitination assays, phosphorylation-site mutants, endogenous protein degradation measurement in HepG2 cells\",\n      \"journal\": \"Proceedings of the National Academy of Sciences of the United States of America\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 — in vitro ubiquitination reconstitution combined with in vivo mutagenesis and pharmacological inhibitors\",\n      \"pmids\": [\"13679577\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2003,\n      \"finding\": \"The androgen receptor physically interacts with FOXO1 (mapped to FOXO1 C-terminus aa 350–655 and AR N-terminal A/B region plus ligand-binding domain) in an AKT-independent manner, blocking FOXO1 DNA binding and suppressing FOXO1-induced Fas ligand expression and apoptosis in prostate cancer cells.\",\n      \"method\": \"In vivo and in vitro co-immunoprecipitation, domain-mapping, FOXO1 AKT-site triple mutant analysis, transcriptional reporter assay\",\n      \"journal\": \"Molecular and cellular biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — reciprocal co-IP with domain mapping plus functional rescue experiments\",\n      \"pmids\": [\"12482965\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2004,\n      \"finding\": \"CK1 phosphorylates FOXO1 at Ser-322 and Ser-325 within its multisite phosphorylation domain in vivo; CK1-mediated phosphorylation of these residues is required for accelerated nuclear exclusion of FOXO1 in response to IGF-1 and insulin, as demonstrated with the selective CK1 inhibitor D4476.\",\n      \"method\": \"Cell-based pharmacological inhibition with D4476, endogenous FOXO1 phosphorylation analysis in H4IIE hepatoma cells\",\n      \"journal\": \"EMBO reports\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — specific site phosphorylation shown with selective inhibitor in cells, single lab\",\n      \"pmids\": [\"14710188\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2005,\n      \"finding\": \"CBP acetylates FOXO1 at Lys-242, Lys-245, and Lys-262, which attenuates its DNA-binding ability; acetylation also increases sensitivity of FOXO1 to PKB-mediated phosphorylation at Ser-253 by reducing DNA-bound protection from kinase access.\",\n      \"method\": \"In vitro kinase assay with DNA competitor, acetylation-mimicking and -deficient mutants, transfection studies\",\n      \"journal\": \"Proceedings of the National Academy of Sciences of the United States of America\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 — in vitro reconstitution with mutagenesis and mechanistic dissection of acetylation-phosphorylation crosstalk\",\n      \"pmids\": [\"16076959\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2005,\n      \"finding\": \"FoxO1 forms a complex with PML and the deacetylase SIRT1 in pancreatic beta cells; acetylation of FoxO1 targets it to PML and prevents ubiquitin-dependent degradation; this complex activates NeuroD and MafA expression to protect beta cells against oxidative stress.\",\n      \"method\": \"Co-immunoprecipitation, acetylation-defective/mimicking mutants, transgenic mouse model expressing constitutively nuclear FoxO1\",\n      \"journal\": \"Cell metabolism\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — Co-IP of endogenous complex plus mutagenesis and in vivo transgenic validation\",\n      \"pmids\": [\"16154098\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2005,\n      \"finding\": \"FoxO1a directly activates transcription of the cGKI (cyclic GMP-dependent protein kinase I) gene during myoblast differentiation; the resulting cGKI then phosphorylates FoxO1a and abolishes its DNA-binding activity, establishing a negative feedback loop that regulates myoblast fusion.\",\n      \"method\": \"Transcriptional reporter assay, in vitro phosphorylation, DNA-binding assay, overexpression studies in myogenic cells\",\n      \"journal\": \"Molecular and cellular biology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — functional feedback loop demonstrated with reporter assay and in vitro phosphorylation, single lab\",\n      \"pmids\": [\"16107711\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2010,\n      \"finding\": \"FoxO1 binds to multiple enhancer-like elements within the Tlr4 gene and Tlr4-pathway genes in macrophages (shown by ChIP-Seq), potentiating TLR4-mediated inflammatory signaling; TLR4 activation in turn induces AKT, which phosphorylates and inactivates FoxO1, creating a self-limiting inflammatory loop.\",\n      \"method\": \"ChIP combined with massively parallel sequencing (ChIP-Seq), genetic FoxO1 manipulation in macrophages\",\n      \"journal\": \"The EMBO journal\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — genome-wide ChIP-Seq plus functional epistasis, multiple orthogonal methods\",\n      \"pmids\": [\"21045807\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2010,\n      \"finding\": \"The TAK1-NLK kinase pathway phosphorylates FOXO1 at Pro-directed Ser/Thr residues in its transactivation domain, inhibiting FOXO1 transcriptional activity and causing nuclear exclusion independently of the PI3K/Akt pathway; NLK directly binds FOXO1 and this interaction is evolutionarily conserved in Drosophila.\",\n      \"method\": \"In vitro kinase assay, co-immunoprecipitation, siRNA knockdown, subcellular localization imaging\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — in vitro kinase assay plus co-IP and functional epistasis, single lab\",\n      \"pmids\": [\"20061393\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2011,\n      \"finding\": \"In beta cells, FoxO1 translocates from cytoplasm to nucleus in response to nitric oxide and activates GADD45α expression to promote DNA repair; SIRT1 deacetylase activity determines whether this response is protective (GADD45α) or proapoptotic (PUMA induction and caspase-3 cleavage).\",\n      \"method\": \"Subcellular fractionation/imaging, SIRT1 inhibitor treatment, siRNA knockdown, mRNA and protein measurements\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — direct localization linked to functional gene expression outcome, single lab\",\n      \"pmids\": [\"21196578\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2003,\n      \"finding\": \"PAK1 directly phosphorylates FOXO1 in a PI3K-independent, estrogen-stimulated manner and physically interacts with FOXO1; PAK1-mediated phosphorylation causes perinuclear cytoplasmic localization of FOXO1 and impairs its ability to activate the Fas ligand promoter.\",\n      \"method\": \"In vitro kinase assay, co-immunoprecipitation, subcellular localization imaging, transcriptional reporter assay\",\n      \"journal\": \"FEBS letters\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — in vitro kinase assay plus co-IP with functional transcriptional readout, single lab\",\n      \"pmids\": [\"12560069\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2004,\n      \"finding\": \"Foxo1-deficient mice die at ~E11 due to severely impaired vascular development; Foxo1-null endothelial cells show abnormal morphological responses to VEGF but not in its absence, demonstrating that Foxo1 is required for proper endothelial cell responses to VEGF during angiogenesis.\",\n      \"method\": \"Knockout mouse generation, embryonic stem cell differentiation assay, VEGF stimulation experiment\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — genetic knockout with defined cellular phenotype and mechanistic VEGF-response assay\",\n      \"pmids\": [\"15184386\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2006,\n      \"finding\": \"FOXO1 binds to the FOXO-responsive element (DBE2) in the Selenoprotein P (SeP) promoter; PGC-1α co-activates this FOXO1-driven expression and cooperates with HNF-4α binding at an adjacent site, coordinately regulating SeP transcription in liver in response to insulin and glucocorticoids.\",\n      \"method\": \"Promoter reporter assay with site-directed mutations, overexpression studies, co-activator interaction analysis\",\n      \"journal\": \"Hepatology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — promoter binding and mutagenesis with functional transcriptional readout, single lab\",\n      \"pmids\": [\"18972406\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2009,\n      \"finding\": \"FOXO1A physically associates with HoxA-11 (demonstrated by co-immunoprecipitation and chromatin immunoprecipitation), and together they bind a decidual prolactin promoter enhancer region; FOXO1A converts HoxA-11 from a repressor to an activator, driving decidualization-specific prolactin expression.\",\n      \"method\": \"Co-immunoprecipitation, chromatin immunoprecipitation, promoter reporter assay, siRNA knockdown\",\n      \"journal\": \"PloS one\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — reciprocal co-IP and ChIP confirming physical complex with functional transcriptional consequence, single lab\",\n      \"pmids\": [\"19727442\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2015,\n      \"finding\": \"FOXO1 directly binds the CCR7 and ICAM-1 promoters (shown by ChIP) and stimulates their transcription in dendritic cells; FOXO1-driven ICAM-1 and CCR7 expression is required for DC phagocytosis, chemotaxis, lymph-node homing, and T/B lymphocyte activation.\",\n      \"method\": \"ChIP, promoter reporter assay, lineage-specific Foxo1 knockout mice, rescue transfection with ICAM-1 or CCR7 plasmids\",\n      \"journal\": \"Journal of immunology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — ChIP plus genetic knockout with rescue experiments and functional in vivo readout\",\n      \"pmids\": [\"25786691\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2015,\n      \"finding\": \"In normal glucose conditions, FOXO1 promotes keratinocyte migration by binding the TGFβ1 promoter and stimulating TGFβ1 expression; in high glucose, FOXO1 nuclear localization is induced but it no longer binds the TGFβ1 promoter, instead driving SERPINB2 and CCL20 expression that inhibits migration.\",\n      \"method\": \"Lineage-specific Foxo1 deletion in mice, ChIP, siRNA knockdown of downstream targets, keratinocyte migration assay\",\n      \"journal\": \"The Journal of cell biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — ChIP with genetic knockout and rescue providing mechanistic pathway placement\",\n      \"pmids\": [\"25918228\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"SCP4 (CTDSPL2) is a nuclear phosphatase that directly dephosphorylates FoxO1 and FoxO3a, promoting nuclear retention and transcriptional activation of gluconeogenic target genes PEPCK1 and G6PC; SCP4 gene ablation causes neonatal hypoglycemia.\",\n      \"method\": \"Ectopic expression and knockdown of SCP4, in vivo phosphatase assay, genetic knockout mouse with hypoglycemia phenotype\",\n      \"journal\": \"Diabetes\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — in vivo phosphatase activity demonstrated with knockout phenotype, single lab\",\n      \"pmids\": [\"28851713\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"DDB1-CUL4A ubiquitin E3 ligase promotes FOXO1 protein stability by degrading the circadian repressor CRY1; in the absence of CRY1, insulin fails to reduce nuclear FOXO1 or suppress gluconeogenic gene expression, placing DDB1-mediated CRY1 degradation upstream of FOXO1 in hepatic glucose regulation.\",\n      \"method\": \"Hepatocyte-specific Ddb1 knockout, Cry1 depletion, protein stability assay, gluconeogenesis gene expression analysis\",\n      \"journal\": \"Diabetes\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — genetic knockouts with mechanistic protein stability assay, single lab\",\n      \"pmids\": [\"28790135\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"FOXO1 directly binds the Il9 and Irf4 promoters (shown by ChIP and luciferase assay) and transactivates them, promoting Th9 cell differentiation and IL-9 production; pharmacological or genetic disruption of Foxo1 in CD4+ T cells reduces IL-9 and shifts cytokine profile toward IL-17A and IFNγ.\",\n      \"method\": \"ChIP, luciferase promoter reporter assay, pharmacological inhibition (AS1842856), CD4-specific Foxo1 knockout, adoptive transfer airway inflammation model\",\n      \"journal\": \"Scientific reports\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — ChIP and reporter assay with genetic and pharmacological validation, single lab\",\n      \"pmids\": [\"29339772\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"Keratinocyte-specific FOXO1 deletion reduces VEGFA expression in wounds, demonstrating that FOXO1 directly regulates VEGFA transcription and is required for endothelial cell proliferation and angiogenesis during wound healing.\",\n      \"method\": \"Lineage-specific Foxo1 knockout mouse, in vitro VEGFA transcription assay, porcine wound model with FOXO1 inhibitor\",\n      \"journal\": \"The Journal of pathology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — genetic knockout with functional angiogenesis phenotype plus in vitro transcription assay, single lab\",\n      \"pmids\": [\"29574902\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"FOXO1 directly binds the KLF5 promoter in cardiomyocytes (shown by ChIP) and increases KLF5 expression; KLF5 in turn binds the NOX4 promoter and induces NOX4 expression, causing oxidative stress and diabetic cardiomyopathy; cardiomyocyte-specific FOXO1 deletion protects mice from this pathway.\",\n      \"method\": \"Cardiomyocyte-specific Foxo1 knockout mice, ChIP, AAV-mediated Klf5 delivery, pharmacological and genetic KLF5 inhibition\",\n      \"journal\": \"Circulation research\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — ChIP plus genetic knockout and rescue with multiple orthogonal methods\",\n      \"pmids\": [\"33539225\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"S100A11 interacts with HDAC6 (co-immunoprecipitation) to inhibit HDAC6 deacetylase activity, leading to increased acetylation and activation of FOXO1, which promotes autophagy and lipogenesis; FOXO1 inhibition reverses S100A11-induced hepatic lipid accumulation.\",\n      \"method\": \"Co-immunoprecipitation, HDAC6 inhibitor treatment, FOXO1 inhibitor treatment, in vitro and in vivo lipid accumulation assays\",\n      \"journal\": \"Cellular and molecular gastroenterology and hepatology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2–3 — co-IP of endogenous proteins with functional pharmacological inhibitor validation, single lab\",\n      \"pmids\": [\"33075563\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"Foxo1 deficiency in intestinal epithelial cells impairs goblet cell autophagy and mucus secretion, leading to dysbiosis and disrupted tight junction organization; Foxo1 thus regulates gut homeostasis by controlling goblet cell function and intestinal barrier integrity.\",\n      \"method\": \"Intestinal epithelial cell-specific Foxo1 knockout mouse, autophagy assay, microbiota analysis, tight junction protein imaging\",\n      \"journal\": \"The Journal of experimental medicine\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — genetic knockout with defined cellular and microbiome phenotypes, multiple orthogonal methods\",\n      \"pmids\": [\"34287641\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"Akt phosphorylates FoxO1 in oligodendrocyte lineage cells; unphosphorylatable FoxO1 (Akt-site mutants) accumulates on the Sox10 promoter and represses it, while Akt-mediated phosphorylation relieves this repression and is required for Sox10 expression and oligodendrocyte maturation.\",\n      \"method\": \"OL-specific Akt triple-conditional knockout mice, Sox10 promoter activity assay, ChIP with FoxO1 mutants\",\n      \"journal\": \"The Journal of neuroscience\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — genetic knockout combined with reporter assay and ChIP with phosphorylation-site mutants\",\n      \"pmids\": [\"34385359\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"Myeloid-specific FoxO1 deletion skews macrophage polarization from M1 to M2 in vivo; mechanistically, FoxO1 counteracts Stat6 to promote M1 over M2 polarization, perpetuating hepatic inflammation in NASH.\",\n      \"method\": \"Myeloid cell conditional Foxo1 knockout mice, macrophage polarization assays, high-fat and NASH-inducing diet models, Stat6 interaction analysis\",\n      \"journal\": \"The Journal of clinical investigation\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — genetic knockout with mechanistic Stat6 antagonism and in vivo disease phenotype, multiple orthogonal methods\",\n      \"pmids\": [\"35700043\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"FOXO1 is a forkhead transcription factor whose activity, subcellular localization, and stability are coordinately regulated by phosphorylation (by Akt/PKB at Thr-24/Ser-256/Ser-319, CK1 at Ser-322/325, DYRK1A at Ser-329, NLK at transactivation-domain residues, and CDK2), acetylation (by CBP, reversed by SIRT1) and ubiquitination; Akt-mediated phosphorylation promotes cytoplasmic sequestration and proteasomal degradation, while nuclear FOXO1 directly binds promoters of gluconeogenic genes (G6Pase, PEPCK), immune regulators (TLR4, CCR7, ICAM-1, IL-9/IRF4), and stress-response genes (GADD45α, KLF5, VEGFA) to coordinate metabolism, cell survival, inflammation, and tissue homeostasis across multiple cell types.\"\n}\n```\n\n--- ROUND 2 (R2/R3, augmented corpus) ---\n\n```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 1999,\n      \"finding\": \"Akt (protein kinase B) directly phosphorylates FOXO1 (FKHR) in vitro and in vivo at three sites (Thr-24, Ser-256, Ser-319), leading to suppression of FOXO1-mediated transcriptional activation; mutation of all three sites to alanine renders FOXO1 resistant to Akt inhibition and causes apoptosis in a DNA-binding-dependent manner.\",\n      \"method\": \"In vitro kinase assay, co-transfection with active/inactive Akt, alanine-substitution mutagenesis, reporter gene assays, apoptosis assay\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 — in vitro phosphorylation plus mutagenesis, independently replicated in two back-to-back papers (PMIDs 10358014 and 10358075)\",\n      \"pmids\": [\"10358014\", \"10358075\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 1999,\n      \"finding\": \"FOXO1 binds in vitro to the insulin-responsive sequence (IRS) in the IGFBP-1 promoter and activates transcription from IRS-containing reporter plasmids; Akt-mediated phosphorylation inhibits this transcriptional activity.\",\n      \"method\": \"In vitro DNA binding assay, luciferase reporter assay, co-transfection\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 — direct DNA-binding assay plus functional reporter, replicated across two concurrent papers\",\n      \"pmids\": [\"10358014\", \"10358075\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2001,\n      \"finding\": \"FOXO1 confers insulin sensitivity onto glucose-6-phosphatase (G6P) expression in kidney epithelial cells; adenoviral transduction of FOXO1 restores insulin-mediated suppression of dexamethasone/cAMP-induced G6P expression, and dominant-negative FOXO1 partially inhibits G6P and PEPCK expression in primary hepatocytes, establishing FOXO1 as a mediator of insulin regulation of gluconeogenic gene expression.\",\n      \"method\": \"Adenoviral transduction, dominant-negative mutant expression, primary hepatocyte cultures, gene expression analysis\",\n      \"journal\": \"The Journal of clinical investigation\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — loss-of-function and gain-of-function with defined molecular phenotype in multiple cell types\",\n      \"pmids\": [\"11696581\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2001,\n      \"finding\": \"DYRK1A phosphorylates FOXO1 (FKHR) specifically at Ser-329 in vitro; this phosphorylation reduces FOXO1 nuclear abundance and transcriptional activity; DYRK1A and FOXO1 co-localize in discrete nuclear regions and can be co-immunoprecipitated, suggesting DYRK1A phosphorylates FOXO1 at Ser-329 in vivo.\",\n      \"method\": \"Purification of kinase from skeletal muscle, in vitro kinase assay, co-immunoprecipitation, nuclear co-localization, site-directed mutagenesis, reporter assay\",\n      \"journal\": \"The Biochemical journal\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1-2 — purified kinase reconstitution, co-IP, and localization data in one study\",\n      \"pmids\": [\"11311120\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2002,\n      \"finding\": \"FOXO1-mediated inhibition of cell cycle progression and transformation requires transcriptional repression of D-type cyclins (Class III targets) rather than activation of insulin-response-sequence genes; chromatin immunoprecipitation confirmed direct FOXO1 occupancy at cyclin D promoters.\",\n      \"method\": \"Transcriptional profiling, chromatin immunoprecipitation (ChIP), functional cell-cycle and transformation assays\",\n      \"journal\": \"Cancer cell\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — ChIP plus functional epistasis with multiple orthogonal methods in one study\",\n      \"pmids\": [\"12150827\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2003,\n      \"finding\": \"Insulin-induced Akt-mediated phosphorylation of FOXO1 targets it for ubiquitination and proteasomal degradation; efficient ubiquitination requires both phosphorylation at the PKB consensus sites and cytoplasmic retention, establishing a two-step negative regulation (nuclear exclusion then proteasomal degradation).\",\n      \"method\": \"Proteasome inhibitor treatment, in vivo and in vitro ubiquitination assays, PI3K pathway inhibitors, phosphorylation-site mutants\",\n      \"journal\": \"Proceedings of the National Academy of Sciences of the United States of America\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1-2 — in vitro ubiquitination reconstitution plus in vivo confirmation with multiple mutants\",\n      \"pmids\": [\"13679577\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2003,\n      \"finding\": \"The androgen receptor (AR) directly interacts with FOXO1 through protein-protein interaction (carboxyl terminus of FOXO1, aa 350–655, with AR A/B region and ligand-binding domain), blocking FOXO1 DNA-binding activity and its ability to induce Fas ligand expression and apoptosis in an AKT-independent manner.\",\n      \"method\": \"In vivo and in vitro co-precipitation, domain-mapping, reporter gene assay, apoptosis/cell-cycle assays\",\n      \"journal\": \"Molecular and cellular biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — reciprocal co-precipitation with domain mapping and functional consequence\",\n      \"pmids\": [\"12482965\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2004,\n      \"finding\": \"CK1 phosphorylates FOXO1a at Ser-322 and Ser-325 within a multisite phosphorylation domain in vivo; pharmacological inhibition of CK1 with D4476 specifically blocks these phosphorylations and attenuates IGF-1/insulin-induced nuclear exclusion of FOXO1a.\",\n      \"method\": \"Cell-permeable CK1 inhibitor (D4476), phospho-specific analysis, nuclear/cytoplasmic fractionation, H4IIE hepatoma cells\",\n      \"journal\": \"EMBO reports\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — site-specific pharmacological inhibition with subcellular localization readout\",\n      \"pmids\": [\"14710188\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2004,\n      \"finding\": \"Foxo1 is essential for normal vascular development; Foxo1-deficient mice die at embryonic day ~11 with severe defects in angiogenesis, and Foxo1-null endothelial cells derived from ES cells show aberrant morphological response to VEGF, indicating Foxo1 is required for proper endothelial cell response to VEGF signaling.\",\n      \"method\": \"Foxo1 knockout mice (embryonic lethal phenotype), ES-cell-derived endothelial cell differentiation assay, VEGF stimulation\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — constitutive knockout with defined vascular phenotype and in vitro mechanistic follow-up\",\n      \"pmids\": [\"15184386\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2004,\n      \"finding\": \"SIRT1 (Sir2) binds and deacetylates FOXO1 at lysine residues (K242, K245, K262) that are acetylated by CBP, reversing the attenuation of FOXO1 transcriptional activity; SIRT1 is recruited to FOXO1 target promoters (MnSOD, p27kip1) and enhances their expression in a deacetylase-activity-dependent manner.\",\n      \"method\": \"Co-immunoprecipitation, in vitro deacetylation assay, ChIP, reporter assay, deacetylase-inactive mutants\",\n      \"journal\": \"Proceedings of the National Academy of Sciences of the United States of America\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1-2 — biochemical reconstitution of deacetylation, ChIP, and catalytic mutants in one study\",\n      \"pmids\": [\"15220471\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2004,\n      \"finding\": \"Foxo transcription factors (including Foxo1) drive atrogin-1/MAFbx ubiquitin ligase expression in skeletal muscle; constitutively active Foxo3 acts on the atrogin-1 promoter to cause transcription and dramatic muscle atrophy; dominant-negative Foxo or RNAi-mediated knockdown prevents atrogin-1 induction and muscle atrophy in vitro and in vivo.\",\n      \"method\": \"Constitutively active Foxo constructs, dominant-negative constructs, RNAi in mouse muscle in vivo, promoter reporter assay, myotube atrophy assay\",\n      \"journal\": \"Cell\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — genetic epistasis with multiple loss- and gain-of-function approaches in vitro and in vivo\",\n      \"pmids\": [\"15109499\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2005,\n      \"finding\": \"FoxO1 protects pancreatic beta cells against oxidative stress by forming a complex with PML and SIRT1 to activate NeuroD and MafA transcription; acetylation of FoxO1 targets it to PML and prevents ubiquitin-dependent degradation, and constitutively nuclear FoxO1 in beta cells prevents hyperglycemia-induced suppression of MafA.\",\n      \"method\": \"Co-immunoprecipitation (FoxO1-PML-SIRT1 complex), acetylation-defective and acetylation-mimicking mutants, transgenic mice, in vivo MafA expression analysis\",\n      \"journal\": \"Cell metabolism\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — complex reconstitution by co-IP, functional mutagenesis, and in vivo transgenic validation\",\n      \"pmids\": [\"16154098\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2005,\n      \"finding\": \"Acetylation of FOXO1 by CBP at Lys-242, Lys-245, and Lys-262 reduces its DNA-binding affinity by neutralizing positive charges required for DNA interaction; additionally, acetylation increases susceptibility of FOXO1 to Akt-mediated phosphorylation at Ser-253, and DNA binding inhibits PKB phosphorylation of FOXO1 in vitro.\",\n      \"method\": \"In vitro DNA binding assays, in vitro kinase assays, acetylation-mimicking and acetylation-deficient mutants, phosphorylation analysis\",\n      \"journal\": \"Proceedings of the National Academy of Sciences of the United States of America\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 — in vitro biochemical reconstitution of acetylation-DNA binding crosstalk with mutagenesis\",\n      \"pmids\": [\"16076959\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2005,\n      \"finding\": \"Skp2, an oncogenic F-box protein, interacts with and ubiquitinates FOXO1 in an Akt phosphorylation (Ser-256)-dependent manner, promoting FOXO1 proteasomal degradation and abolishing its growth-inhibitory and pro-apoptotic functions.\",\n      \"method\": \"Co-immunoprecipitation, in vitro ubiquitination assay, phosphorylation-site mutants, cell proliferation/survival assays, mouse lymphoma model\",\n      \"journal\": \"Proceedings of the National Academy of Sciences of the United States of America\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1-2 — biochemical reconstitution of ubiquitination with site-specific mutants and in vivo model\",\n      \"pmids\": [\"15668399\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2005,\n      \"finding\": \"Foxo1 (and Foxo3a) overexpression inhibits endothelial cell migration and tube formation; Foxo1 specifically represses angiopoietin-2 expression and binds to the eNOS promoter to repress eNOS expression, establishing Foxo1 as a regulator of angiogenesis-related gene expression in endothelial cells.\",\n      \"method\": \"Constitutively active Foxo constructs, siRNA knockdown, gene expression profiling, ChIP on eNOS promoter, migration/tube formation assays\",\n      \"journal\": \"The Journal of clinical investigation\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — ChIP plus gain/loss-of-function with defined molecular and cellular phenotypes\",\n      \"pmids\": [\"16100571\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2005,\n      \"finding\": \"PAK1 (p21-activated kinase 1) directly interacts with FOXO1 (FKHR) and phosphorylates it in response to estrogen stimulation, causing perinuclear cytoplasmic retention and preventing FOXO1 from activating Fas ligand promoter transcription; dominant-negative PAK1 restores FOXO1 transcriptional activity.\",\n      \"method\": \"Co-immunoprecipitation, in vitro kinase assay (PAK1 phosphorylating FKHR), localization microscopy, reporter gene assay\",\n      \"journal\": \"FEBS letters\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — direct kinase assay and co-IP, single lab\",\n      \"pmids\": [\"12560069\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2005,\n      \"finding\": \"Beta-catenin directly binds FOXO transcription factors and enhances FOXO transcriptional activity in mammalian cells; this interaction is evolutionarily conserved (C. elegans BAR-1/DAF-16) and is enhanced under oxidative stress, establishing beta-catenin as a co-activator of FOXO-dependent stress response.\",\n      \"method\": \"Co-immunoprecipitation, reporter assay, C. elegans genetic epistasis, oxidative stress experiments\",\n      \"journal\": \"Science\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — direct binding demonstrated by co-IP, replicated across mammalian cells and C. elegans\",\n      \"pmids\": [\"15905404\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2005,\n      \"finding\": \"FoxO1a directly activates transcription of the cyclic GMP-dependent protein kinase I (cGKI) gene; the resulting cGKI then phosphorylates FoxO1a and abolishes its DNA-binding activity, establishing an autoregulatory feedback loop that controls myoblast fusion.\",\n      \"method\": \"Reporter gene assay, phosphorylation assay (cGKI on FoxO1a), myoblast fusion assay\",\n      \"journal\": \"Molecular and cellular biology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — direct kinase assay plus functional consequence in differentiation, single lab\",\n      \"pmids\": [\"16107711\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2006,\n      \"finding\": \"Constitutively active FoxO1 in transgenic mouse liver increases gluconeogenic gene expression (PEPCK, G6Pase), suppresses glycolytic, pentose phosphate shunt, and lipogenic genes (including glucokinase and SREBP-1c), and impairs glucose tolerance; adenoviral FoxO1 in hepatocytes confirms direct regulation of these metabolic gene programs.\",\n      \"method\": \"Transgenic mice with liver-specific constitutively active FoxO1, gene array, adenoviral transduction of hepatocytes, metabolic phenotyping\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — in vivo transgenic and in vitro adenoviral models with genome-wide gene expression analysis\",\n      \"pmids\": [\"16492665\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2006,\n      \"finding\": \"FOXO1A cooperates with HoxA-10 to regulate decidual IGFBP-1 expression; co-immunoprecipitation and ChIP demonstrate physical association of FOXO1A with HoxA-11 and binding of both to the decidual prolactin promoter enhancer, forming a decidual-specific transcriptional complex that switches HoxA-11 from repressor to activator.\",\n      \"method\": \"Co-immunoprecipitation, ChIP, reporter gene assay, siRNA knockdown\",\n      \"journal\": \"PloS one\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — co-IP and ChIP with functional reporter assay, single lab\",\n      \"pmids\": [\"19727442\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2008,\n      \"finding\": \"PRMT1 methylates FOXO1 at Arg-248 and Arg-250 within the Akt phosphorylation consensus motif; this arginine methylation directly blocks Akt-mediated phosphorylation of FOXO1 at Ser-253 in vitro and in vivo, preventing nuclear exclusion and proteasomal degradation, thereby promoting oxidative-stress-induced apoptosis.\",\n      \"method\": \"In vitro methylation assay, in vitro kinase assay, PRMT1 siRNA knockdown, polyubiquitination assay, apoptosis assay, phosphorylation-deficient mutants\",\n      \"journal\": \"Molecular cell\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 — in vitro biochemical reconstitution of methylation-phosphorylation crosstalk with mutagenesis, multiple orthogonal methods\",\n      \"pmids\": [\"18951090\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2008,\n      \"finding\": \"PGC-1alpha cooperates with FoxO1a and HNF-4alpha to drive selenoprotein P (SeP) promoter activity; PGC-1alpha enhances FoxO1a-stimulated SeP transcription through a FoxO-responsive element; insulin suppresses this axis, establishing FoxO1a as a component of the PGC-1alpha-regulated selenium homeostasis network.\",\n      \"method\": \"Reporter assay, promoter mutagenesis, adenoviral overexpression, primary rat hepatocytes, mRNA/protein analysis\",\n      \"journal\": \"Hepatology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — promoter mutagenesis and overexpression in primary cells, single lab\",\n      \"pmids\": [\"18972406\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2010,\n      \"finding\": \"FoxO1 promotes inflammation by enhancing TLR4-mediated signaling in macrophages; ChIP-Seq shows FoxO1 binds multiple enhancer-like elements within the Tlr4 gene and TLR4 pathway genes; TLR4 activation induces Akt which inactivates FoxO1, establishing a self-limiting inflammatory feedback loop.\",\n      \"method\": \"ChIP combined with massively parallel sequencing (ChIP-Seq), FoxO1 knockout macrophages, reporter assay, inflammatory cytokine measurements\",\n      \"journal\": \"The EMBO journal\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — genome-wide ChIP-Seq plus loss-of-function in primary macrophages, multiple orthogonal methods\",\n      \"pmids\": [\"21045807\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2010,\n      \"finding\": \"TAK1-NLK (Nemo-like kinase) pathway phosphorylates FOXO1 at Pro-directed Ser/Thr residues in the transactivation domain; this phosphorylation inhibits FOXO1 transcriptional activity and causes nuclear exclusion independently of PI3K/Akt; NLK binds FOXO1 and this interaction is evolutionarily conserved in Drosophila.\",\n      \"method\": \"Co-immunoprecipitation, kinase assay (NLK on FOXO1), nuclear/cytoplasmic fractionation, siRNA knockdown of TAK1/NLK, FOXO target gene mRNA assay, PARP cleavage\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — direct binding and kinase assay with functional localization data, single lab\",\n      \"pmids\": [\"20061393\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2010,\n      \"finding\": \"Cytosolic FoxO1 mediates autophagy independently of its transcriptional activity; in response to oxidative stress or serum starvation, FoxO1 is acetylated by dissociation from SIRT2, and acetylated FoxO1 binds to Atg7 (an E1-like autophagy protein) to induce autophagy and cell death; this is associated with tumor suppressor activity.\",\n      \"method\": \"Co-immunoprecipitation (FoxO1-Atg7 interaction), transcription-deficient FoxO1 mutants, autophagy assays, SIRT2 interaction studies, xenograft tumor model\",\n      \"journal\": \"Nature cell biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — direct protein-protein interaction by co-IP, functional mutants separating transcriptional from cytoplasmic role, in vivo tumor model\",\n      \"pmids\": [\"20543840\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2011,\n      \"finding\": \"FoxO1 and SIRT1 regulate beta-cell responses to nitric oxide; FoxO1 translocates from cytoplasm to nucleus in response to nitric oxide and stimulates GADD45α expression for DNA repair; SIRT1 regulates this process—SIRT1 inhibition shifts FoxO1 from a protective (GADD45α/DNA repair) to a proapoptotic (PUMA/caspase-3) program.\",\n      \"method\": \"Nuclear translocation assay, gene expression analysis, SIRT1 pharmacological inhibitors, siRNA, caspase-3 cleavage assay\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — direct localization experiments with functional consequence in primary beta cells, single lab\",\n      \"pmids\": [\"21196578\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2011,\n      \"finding\": \"Hepatic deletion of Foxo1 (but not Foxo3 or Foxo4 alone) reduces blood glucose by decreasing gluconeogenic enzyme gene expression; combined deletion of Foxo1 and Foxo3 additionally increases hepatic lipid synthesis and secretion, demonstrating that Foxo1 specifically controls gluconeogenesis while Foxo3 regulates lipogenic gene expression.\",\n      \"method\": \"Liver-specific Cre/LoxP conditional knockout mice (individual and combined deletions), metabolic phenotyping, liver transcript analysis\",\n      \"journal\": \"Endocrinology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — conditional knockout with genetic epistasis distinguishing paralog-specific functions in vivo\",\n      \"pmids\": [\"22147007\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2013,\n      \"finding\": \"mTORC2 promotes inactivating phosphorylation of class IIa HDACs, leading to acetylation of FoxO1 (and FoxO3), which releases c-Myc from a suppressive miR-34c network to drive glycolytic reprogramming in glioblastoma; this is an Akt-independent mechanism linking mTORC2 to FoxO acetylation and cancer metabolism.\",\n      \"method\": \"mTORC2 genetic manipulation, HDAC phosphorylation/acetylation assays, FoxO acetylation analysis, c-Myc/miR-34c network analysis, glioblastoma patient samples\",\n      \"journal\": \"Cell metabolism\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — mechanistic epistasis across multiple nodes with clinical correlation, multiple orthogonal methods\",\n      \"pmids\": [\"24140020\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"FOXO1/3 knockdown in vascular smooth muscle cells phenocopies PTEN deficiency by increasing Runx2 protein via decreased Runx2 ubiquitination, promoting vascular calcification; AKT activation causes FOXO1/3 nuclear exclusion, establishing a PTEN/AKT/FOXO1/3/Runx2 ubiquitination axis controlling VSMC calcification.\",\n      \"method\": \"SMC-specific PTEN knockout mice (Cre/LoxP), siRNA knockdown, in vitro calcification assay, Runx2 ubiquitination assay, ex vivo arterial calcification\",\n      \"journal\": \"Arteriosclerosis, thrombosis, and vascular biology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — in vivo genetic model plus in vitro mechanistic dissection, single lab\",\n      \"pmids\": [\"25378413\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2015,\n      \"finding\": \"FOXO1 directly regulates CCR7 and ICAM-1 promoters (shown by ChIP and promoter reporter assays) in dendritic cells; FOXO1 nuclear localization is induced by bacteria via the MAPK pathway; lineage-specific FOXO1 deletion impairs DC phagocytosis, chemotaxis, and lymph node homing, which can be rescued by ICAM-1 and CCR7 re-expression.\",\n      \"method\": \"ChIP, promoter reporter assay, lineage-specific knockout (CD11c.Cre), rescue with plasmid transfection, in vivo lymph node homing assay\",\n      \"journal\": \"Journal of immunology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — ChIP plus genetic rescue in vivo demonstrating direct target gene mechanism\",\n      \"pmids\": [\"25786691\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2015,\n      \"finding\": \"FOXO1 enhances keratinocyte migration and wound healing by upregulating TGFβ1 under normal glucose conditions; under high glucose, FOXO1 nuclear localization is induced but FOXO1 does not bind the TGFβ1 promoter—instead it drives SERPINB2 and CCL20 expression that inhibits migration; insulin treatment or FOXO1 silencing rescues impaired migration in high glucose.\",\n      \"method\": \"Lineage-specific Foxo1 deletion, FOXO1 promoter binding analysis, siRNA, high-glucose cell culture, wound healing migration assay\",\n      \"journal\": \"The Journal of cell biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — in vivo conditional knockout with mechanistic promoter binding and rescue experiments\",\n      \"pmids\": [\"25918228\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"FOXO1 suppresses MYC signaling in endothelial cells to restrain glycolysis and mitochondrial respiration, acting as a gatekeeper of endothelial quiescence; endothelial-restricted FOXO1 deletion causes increased EC proliferation and vessel hyperplasia, while forced FOXO1 expression restricts vascular expansion; restoration of MYC in FOXO1-overexpressing endothelium normalizes metabolism and branching.\",\n      \"method\": \"Endothelial-specific knockout and overexpression mouse models, metabolic flux assays (glycolysis, respiration), MYC rescue experiments, EC proliferation assays\",\n      \"journal\": \"Nature\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — in vivo endothelial-specific genetic models with mechanistic epistasis and metabolic phenotyping, multiple orthogonal approaches\",\n      \"pmids\": [\"26735015\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"DDB1-CUL4A E3 ubiquitin ligase promotes FOXO1-driven hepatic gluconeogenesis by degrading CRY1 (a known DDB1 target); loss of Ddb1 in hepatocytes downregulates FOXO1 protein and impairs gluconeogenic response; CRY1 depletion increases nuclear FOXO1 abundance and enhances gluconeogenesis, linking circadian clock protein degradation to FOXO1 stability.\",\n      \"method\": \"Hepatocyte-specific Ddb1 knockout mice, Cry1 depletion in vivo, FOXO1 protein stability analysis, gluconeogenic gene expression, glucose production assay\",\n      \"journal\": \"Diabetes\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — conditional knockout with mechanistic pathway dissection, single lab\",\n      \"pmids\": [\"28790135\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"SCP4 (CTDSPL2) is a nuclear phosphatase that directly dephosphorylates FoxO1/3a, promoting nuclear retention and transcriptional activation of PEPCK1 and G6PC gluconeogenic genes; SCP4 overexpression increases hepatic glucose production; SCP4 gene ablation causes neonatal hypoglycemia.\",\n      \"method\": \"In vitro dephosphorylation assay (SCP4 on FoxO1/3a), ectopic expression, siRNA knockdown, SCP4 knockout mice (neonatal hypoglycemia), glucose production assay\",\n      \"journal\": \"Diabetes\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1-2 — direct in vitro dephosphorylation reconstitution plus in vivo knockout phenotype\",\n      \"pmids\": [\"28851713\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"FOXO1 directly regulates VEGFA expression in keratinocytes (shown by in vitro promoter assay); keratinocyte-specific FOXO1 deletion reduces VEGFA expression in wounds, resulting in reduced endothelial cell proliferation, reduced angiogenesis, and impaired wound re-epithelialization.\",\n      \"method\": \"Keratinocyte-specific Foxo1 knockout mice, VEGFA promoter reporter assay, endothelial proliferation assay, porcine wound model with FOXO1 inhibitor\",\n      \"journal\": \"The Journal of pathology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — in vivo conditional knockout with direct promoter binding evidence and large-animal model\",\n      \"pmids\": [\"29574902\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"Foxo1 directly binds the Il9 and Irf4 promoters in Th9 cells (shown by ChIP and luciferase assays) to transactivate them; pharmacological or genetic disruption of Foxo1 in CD4+ T cells reduces IL-9 and promotes IL-17A/IFNγ; adoptive transfer of Foxo1-inhibitor-treated Th9 cells reduces asthma-like symptoms.\",\n      \"method\": \"ChIP, luciferase promoter assay, Foxo1 pharmacological inhibitor (AS1842856), CD4-specific Foxo1 knockout, adoptive transfer airway allergy model\",\n      \"journal\": \"Scientific reports\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — ChIP plus loss-of-function with defined cellular phenotype, single lab\",\n      \"pmids\": [\"29339772\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"Glucagon suppresses hepatic mitochondrial complex III and IV activity and reduces mitochondrial biogenesis (NRF1, TFAM, MFN2 expression) in a FOXO1-dependent manner; hepatocyte-specific Foxo1 deletion largely rescues glucagon-induced mitochondrial dysfunction, establishing FOXO1 as a mediator of glucagon's effects on mitochondrial function.\",\n      \"method\": \"Foxo1 hepatocyte-specific knockout mice, glucagon treatment, mitochondrial function assays (complex activity), db/db diabetic mice\",\n      \"journal\": \"The Journal of endocrinology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — conditional knockout with functional mitochondrial phenotype, single lab\",\n      \"pmids\": [\"31026811\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"METTL14 promotes FOXO1 expression in endothelial cells by directly binding FOXO1 mRNA, increasing its m6A modification, and enhancing translation through YTHDF1 recognition; METTL14 also interacts with FOXO1 protein and together they act on VCAM-1 and ICAM-1 promoters to promote inflammatory gene transcription.\",\n      \"method\": \"Methylated RNA immunoprecipitation (RIP) sequencing, RIP assay, co-immunoprecipitation, ChIP, METTL14 knockout mice, TNF-α inflammation model\",\n      \"journal\": \"Theranostics\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — RIP-seq, co-IP, and ChIP with in vivo knockout, single lab\",\n      \"pmids\": [\"32802173\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"FOXO1 binds directly to the KLF5 promoter in cardiomyocytes to increase KLF5 expression; KLF5 in turn binds the NOX4 promoter to induce NOX4 expression, reactive oxygen species, and ceramide accumulation, causing diabetic cardiomyopathy; cardiomyocyte-specific FOXO1 deletion protects from diabetic cardiomyopathy by reducing KLF5.\",\n      \"method\": \"ChIP (FOXO1 on KLF5 promoter; KLF5 on NOX4 promoter), cardiomyocyte-specific FOXO1 knockout mice, AAV-mediated KLF5 delivery, pharmacological KLF5 inhibition, oxidative stress and ceramide measurements\",\n      \"journal\": \"Circulation research\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — ChIP-validated direct target gene regulation, in vivo knockout, and multiple orthogonal rescue approaches\",\n      \"pmids\": [\"33539225\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"S100A11 interacts with HDAC6 to inhibit HDAC6 deacetylase activity, thereby releasing FOXO1 from HDAC6 binding and increasing FOXO1 acetylation; acetylated FOXO1 activates autophagy and lipogenesis programs promoting liver steatosis.\",\n      \"method\": \"Co-immunoprecipitation (S100A11-HDAC6 interaction), HDAC6 inhibitor, FOXO1 inhibitor, adenoviral overexpression, in vivo high-fat diet model, lipid staining\",\n      \"journal\": \"Cellular and molecular gastroenterology and hepatology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — co-IP with pharmacological and genetic dissection in vitro and in vivo, single lab\",\n      \"pmids\": [\"33075563\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"Akt phosphorylation of FoxO1 is required for Sox10 expression in oligodendrocytes; unphosphorylated (nuclear) FoxO1 represses the Sox10 promoter; Akt-triple-knockout oligodendrocyte lineage mice lack mature oligodendrocytes and myelination with downregulated Sox10, identifying a FoxO1 phosphorylation-dependent mechanism for oligodendrocyte differentiation.\",\n      \"method\": \"OL lineage-specific Akt1/2/3 triple conditional knockout mice, Sox10 promoter reporter assay, FoxO1 phosphorylation-site mutants, ChIP (FoxO1 on Sox10 promoter)\",\n      \"journal\": \"The Journal of neuroscience\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — in vivo triple knockout with direct promoter occupancy evidence, single lab\",\n      \"pmids\": [\"34385359\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"Foxo1 in intestinal epithelial cells regulates goblet cell autophagy and mucus secretion; IEC-specific Foxo1 deletion impairs goblet cell autophagy and mucus secretion, leading to gut microbiota dysbiosis, disrupted tight junction organization, and increased susceptibility to intestinal inflammation.\",\n      \"method\": \"Intestinal epithelial cell-specific Foxo1 conditional knockout mice, goblet cell autophagy assays, microbiome analysis, tight junction protein analysis, colitis susceptibility assay\",\n      \"journal\": \"The Journal of experimental medicine\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — in vivo conditional knockout with defined cellular mechanism (autophagy-mucus axis), single lab\",\n      \"pmids\": [\"34287641\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"FoxO1 in hepatic macrophages (Kupffer cells) promotes pro-inflammatory M1 polarization by counteracting Stat6 signaling; myeloid-specific FoxO1 knockout skews macrophage polarization from M1 to M2 and reduces hepatic inflammation, steatosis, and fibrosis in NASH.\",\n      \"method\": \"Myeloid-specific FoxO1 conditional knockout (LysM-Cre), M1/M2 polarization markers, Stat6 pathway analysis, NASH diet model, high-fat diet model\",\n      \"journal\": \"The Journal of clinical investigation\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — in vivo conditional knockout with mechanistic epistasis (FoxO1-Stat6) and disease-relevant phenotype\",\n      \"pmids\": [\"35700043\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"FOXO1 directly binds the promoter regions of CD36 and STEAP4 genes in bovine adipocytes (shown by ChIP and site-directed mutagenesis) and regulates their transcriptional activity, linking FOXO1 to lipogenesis through the AMPK and PI3K/AKT pathways.\",\n      \"method\": \"ChIP, site-directed mutagenesis of FOXO1 binding sites, transcriptomic and metabolomics analyses, siRNA knockdown\",\n      \"journal\": \"International journal of biological macromolecules\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — ChIP plus promoter mutagenesis, single lab, non-human model\",\n      \"pmids\": [\"37506793\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"FOXO1 is a forkhead-family transcription factor that is negatively regulated by Akt-mediated phosphorylation (at Thr-24, Ser-256, Ser-319), which drives its nuclear exclusion, ubiquitination by Skp2, and proteasomal degradation; its activity is further modulated by CK1- and DYRK1A-mediated phosphorylation, CBP-mediated acetylation (attenuating DNA binding and increasing Akt sensitivity), SIRT1/SIRT2-mediated deacetylation (enhancing transcriptional activity or autophagy), PRMT1-mediated arginine methylation (blocking Akt phosphorylation), and dephosphorylation by SCP4; in the nucleus, FOXO1 binds insulin-response sequences to activate gluconeogenic genes (G6Pase, PEPCK) and pro-apoptotic targets (FasL, PUMA, GADD45α), represses D-type cyclins and MYC to enforce cell-cycle arrest and metabolic quiescence, and forms functional complexes with PML-SIRT1, beta-catenin, Smad proteins, HoxA factors, and Atg7 (in a transcription-independent autophagy pathway) to integrate insulin/PI3K, oxidative stress, TGFβ, and Wnt signals across diverse cell types including hepatocytes, endothelial cells, macrophages, beta cells, and T cells.\"\n}\n```","stage2_raw":"```json\n{\n  \"mechanistic_narrative\": \"FOXO1 is a forkhead-family transcription factor that integrates insulin/PI3K/Akt signaling with transcriptional programs controlling gluconeogenesis, cell survival, immune polarization, angiogenesis, and autophagy across diverse cell types. Akt phosphorylates FOXO1 at Thr-24, Ser-256, and Ser-319 to drive its cytoplasmic sequestration, ubiquitination, and proteasomal degradation, while additional kinases (CK1, DYRK1A, NLK, PAK1) and the phosphatase SCP4 fine-tune nuclear residence independently of PI3K [PMID:10358014, PMID:14710188, PMID:11311120, PMID:28851713]. CBP-mediated acetylation at Lys-242/245/262 attenuates DNA binding and sensitizes FOXO1 to Akt phosphorylation, whereas SIRT1-dependent deacetylation restores transcriptional competence and directs stress-protective gene programs [PMID:16076959, PMID:16154098]. Nuclear FOXO1 directly occupies promoters of metabolic genes (G6Pase, PEPCK), immune regulators (TLR4, CCR7, IL-9/IRF4), and tissue-remodeling factors (VEGFA, KLF5, TGFβ1), and cell-type-specific knockouts demonstrate essential roles in hepatic glucose output, macrophage M1 polarization, dendritic-cell homing, goblet-cell autophagy, oligodendrocyte maturation, and embryonic angiogenesis [PMID:11696581, PMID:21045807, PMID:25786691, PMID:34287641, PMID:34385359, PMID:15184386].\",\n  \"teleology\": [\n    {\n      \"year\": 1999,\n      \"claim\": \"Establishing that FOXO1 is a direct Akt substrate resolved how insulin/PI3K signaling silences a forkhead transcription factor: Akt phosphorylation at Thr-24/Ser-256/Ser-319 promotes cytoplasmic retention and blocks transcriptional activity.\",\n      \"evidence\": \"In vitro kinase assays, site-directed mutagenesis, and transcriptional reporters in 293T cells with pharmacological pathway epistasis\",\n      \"pmids\": [\"10358014\", \"10358075\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\n        \"Mechanism of nuclear export after phosphorylation not molecularly defined\",\n        \"Whether phosphorylation leads to degradation was unknown at this point\",\n        \"The identity of FOXO1 target genes in physiological contexts remained open\"\n      ]\n    },\n    {\n      \"year\": 2001,\n      \"claim\": \"Identifying G6Pase and PEPCK as FOXO1-dependent insulin-responsive genes established FOXO1 as the key transcription factor mediating insulin suppression of hepatic gluconeogenesis.\",\n      \"evidence\": \"Adenoviral dominant-negative FOXO1 in primary hepatocytes and kidney cells with reporter assays\",\n      \"pmids\": [\"11696581\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\n        \"Genome-wide scope of hepatic FOXO1 targets not yet determined\",\n        \"Whether FOXO1 cooperates with co-activators at gluconeogenic promoters was unknown\"\n      ]\n    },\n    {\n      \"year\": 2001,\n      \"claim\": \"Discovery that DYRK1A phosphorylates FOXO1 at Ser-329 independently of PI3K/Akt revealed a parallel kinase input that reduces nuclear FOXO1 abundance.\",\n      \"evidence\": \"Biochemical kinase purification, in vitro kinase assay, co-IP and co-localization\",\n      \"pmids\": [\"11311120\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\n        \"Physiological stimulus activating DYRK1A toward FOXO1 not identified\",\n        \"No in vivo genetic confirmation of this regulatory axis\"\n      ]\n    },\n    {\n      \"year\": 2002,\n      \"claim\": \"ChIP-confirmed FOXO1 occupancy at cyclin D loci showed that FOXO1 suppresses cell cycle progression through transcriptional repression of D-type cyclins, linking it to tumor suppression.\",\n      \"evidence\": \"Transcriptional profiling, ChIP, and transformation assays\",\n      \"pmids\": [\"12150827\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\n        \"Whether cyclin D repression involves co-repressor recruitment was not determined\",\n        \"Relevance to specific cancer types in vivo not yet tested\"\n      ]\n    },\n    {\n      \"year\": 2003,\n      \"claim\": \"Demonstrating that Akt-phosphorylated FOXO1 is ubiquitinated and degraded by the proteasome added a degradation layer beyond cytoplasmic sequestration, explaining how prolonged insulin signaling depletes FOXO1 protein.\",\n      \"evidence\": \"In vivo/in vitro ubiquitination assays with phospho-site mutants and proteasome inhibitors in HepG2 cells\",\n      \"pmids\": [\"13679577\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\n        \"Identity of the E3 ubiquitin ligase responsible was not determined\",\n        \"Kinetics of degradation versus re-import into nucleus not resolved\"\n      ]\n    },\n    {\n      \"year\": 2003,\n      \"claim\": \"Physical interaction between androgen receptor and FOXO1 that blocks FOXO1 DNA binding revealed a PI3K/Akt-independent mechanism for suppressing FOXO1 pro-apoptotic activity in prostate cancer cells.\",\n      \"evidence\": \"Reciprocal co-IP, domain mapping, and transcriptional reporter with Akt-site triple-mutant FOXO1\",\n      \"pmids\": [\"12482965\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\n        \"Whether AR-FOXO1 interaction occurs genome-wide or at specific loci was unknown\",\n        \"In vivo relevance to prostate cancer progression not tested\"\n      ]\n    },\n    {\n      \"year\": 2004,\n      \"claim\": \"CK1 phosphorylation at Ser-322/325 was shown to accelerate FOXO1 nuclear exclusion after IGF-1 stimulation, revealing a sequential multi-kinase phosphorylation cascade (Akt then CK1) for efficient cytoplasmic retention.\",\n      \"evidence\": \"Selective CK1 inhibitor D4476 in H4IIE hepatoma cells with endogenous FOXO1 phosphorylation analysis\",\n      \"pmids\": [\"14710188\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"Whether CK1 phosphorylation requires prior Akt priming was not formally demonstrated\",\n        \"Single-lab finding; independent replication needed\"\n      ]\n    },\n    {\n      \"year\": 2004,\n      \"claim\": \"Foxo1 knockout embryonic lethality (~E11) from vascular defects established FOXO1 as essential for endothelial responses to VEGF during angiogenesis.\",\n      \"evidence\": \"Foxo1-null mice and ES-cell-derived endothelial differentiation with VEGF stimulation\",\n      \"pmids\": [\"15184386\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\n        \"Direct transcriptional targets mediating the vascular phenotype not identified at this stage\",\n        \"Redundancy with FOXO3/FOXO4 in endothelium not addressed\"\n      ]\n    },\n    {\n      \"year\": 2005,\n      \"claim\": \"CBP acetylation of FOXO1 at Lys-242/245/262 was shown to attenuate DNA binding and increase susceptibility to Akt phosphorylation, while SIRT1 deacetylation in a PML-body complex restores activity, establishing acetylation as a key regulatory switch that cross-talks with phosphorylation.\",\n      \"evidence\": \"In vitro reconstitution with acetylation-mimicking mutants, co-IP of endogenous PML-SIRT1-FOXO1 complex, transgenic mouse beta-cell model\",\n      \"pmids\": [\"16076959\", \"16154098\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\n        \"Whether acetylation globally redirects FOXO1 target gene selection was unknown\",\n        \"Structural basis of how acetylation weakens forkhead-DNA contacts not resolved\"\n      ]\n    },\n    {\n      \"year\": 2010,\n      \"claim\": \"ChIP-Seq in macrophages revealed FOXO1 occupancy at TLR4 and TLR4-pathway gene enhancers, with TLR4 activating Akt to inactivate FOXO1 — establishing a self-limiting inflammatory feedback loop and expanding FOXO1's role to innate immunity.\",\n      \"evidence\": \"ChIP-Seq in macrophages with genetic FOXO1 manipulation\",\n      \"pmids\": [\"21045807\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\n        \"Whether FOXO1 cooperates with NF-κB at these inflammatory loci was not resolved\",\n        \"Contribution to systemic inflammatory disease not directly tested\"\n      ]\n    },\n    {\n      \"year\": 2015,\n      \"claim\": \"FOXO1 ChIP at CCR7 and ICAM-1 promoters in dendritic cells, combined with lineage-specific knockout and rescue, demonstrated that FOXO1 drives DC homing and antigen presentation capacity, broadening its immune role beyond macrophages.\",\n      \"evidence\": \"ChIP, reporter assays, DC-specific Foxo1 knockout mice with plasmid rescue\",\n      \"pmids\": [\"25786691\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\n        \"Whether FOXO1 controls additional DC-specific gene programs genome-wide was not determined\",\n        \"Interaction with other forkhead family members in DCs not assessed\"\n      ]\n    },\n    {\n      \"year\": 2017,\n      \"claim\": \"Identification of the phosphatase SCP4 as a direct FOXO1 dephosphorylating enzyme that promotes nuclear retention and gluconeogenic gene activation — with knockout causing neonatal hypoglycemia — revealed the 'off-switch' counterpart to Akt-mediated inactivation.\",\n      \"evidence\": \"In vivo phosphatase assay, SCP4 knockout mice with hypoglycemia phenotype\",\n      \"pmids\": [\"28851713\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"Whether SCP4 acts on all three Akt sites equally was not determined\",\n        \"Regulation of SCP4 itself in metabolic contexts is unclear\"\n      ]\n    },\n    {\n      \"year\": 2020,\n      \"claim\": \"FOXO1 ChIP at the KLF5 promoter in cardiomyocytes linked FOXO1 to oxidative stress via a FOXO1→KLF5→NOX4 transcriptional cascade causing diabetic cardiomyopathy, revealing a tissue-specific pathological output of FOXO1 activity.\",\n      \"evidence\": \"Cardiomyocyte-specific Foxo1 knockout, ChIP, AAV-mediated rescue\",\n      \"pmids\": [\"33539225\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\n        \"Whether FOXO1 activates KLF5 in non-cardiac tissues is unknown\",\n        \"Direct mechanism of FOXO1 recruitment to KLF5 promoter not structurally resolved\"\n      ]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"Conditional knockouts in intestinal epithelium and oligodendrocyte lineage demonstrated that FOXO1 controls goblet cell autophagy/barrier integrity and represses Sox10 to gate oligodendrocyte maturation, extending its role to gut homeostasis and CNS myelination.\",\n      \"evidence\": \"Intestinal epithelial Foxo1 knockout with microbiota and autophagy analysis; OL-specific Akt knockout with FoxO1 mutant ChIP on Sox10 promoter\",\n      \"pmids\": [\"34287641\", \"34385359\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\n        \"Whether FOXO1's gut barrier role involves direct transcriptional targets on autophagy genes or indirect signaling is not resolved\",\n        \"Redundancy with other FOXO family members in oligodendrocytes not tested\"\n      ]\n    },\n    {\n      \"year\": 2022,\n      \"claim\": \"Myeloid-specific FoxO1 deletion shifted macrophages from M1 to M2 by relieving FOXO1's antagonism of Stat6, identifying FOXO1 as a polarization switch that drives hepatic inflammation in NASH.\",\n      \"evidence\": \"Myeloid conditional Foxo1 knockout on NASH diet with Stat6 interaction analysis\",\n      \"pmids\": [\"35700043\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\n        \"Molecular mechanism of FOXO1-Stat6 antagonism (competitive binding, co-repression, or sequestration) not defined\",\n        \"Whether pharmacological FOXO1 inhibition in macrophages is therapeutic for NASH not tested\"\n      ]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"Key unresolved questions include the identity of the E3 ubiquitin ligase(s) that directly ubiquitinate phospho-FOXO1, the structural basis for how acetylation and phosphorylation combinatorially reprogram FOXO1 target gene selection, and whether FOXO1's diverse tissue-specific outputs reflect distinct co-factor complexes or chromatin accessibility states.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Low\",\n      \"gaps\": [\n        \"E3 ligase directly ubiquitinating FOXO1 not identified\",\n        \"No structural model of post-translationally modified FOXO1 on DNA\",\n        \"Chromatin context determining tissue-specific target selection unknown\"\n      ]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0003677\", \"supporting_discovery_ids\": [2, 4, 11, 17, 18, 19, 22, 24, 27]},\n      {\"term_id\": \"GO:0140110\", \"supporting_discovery_ids\": [0, 2, 4, 11, 16, 17, 18, 19, 22, 24, 27, 28]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005634\", \"supporting_discovery_ids\": [0, 3, 7, 8, 9, 13, 19, 20, 27]},\n      {\"term_id\": \"GO:0005829\", \"supporting_discovery_ids\": [0, 5, 7, 12, 14]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-162582\", \"supporting_discovery_ids\": [0, 1, 5, 7, 12, 14, 20]},\n      {\"term_id\": \"R-HSA-74160\", \"supporting_discovery_ids\": [2, 4, 11, 16, 17, 18, 19, 22, 24, 27]},\n      {\"term_id\": \"R-HSA-1430728\", \"supporting_discovery_ids\": [2, 16, 20, 21]},\n      {\"term_id\": \"R-HSA-168256\", \"supporting_discovery_ids\": [11, 18, 22, 28]},\n      {\"term_id\": \"R-HSA-9612973\", \"supporting_discovery_ids\": [25, 26]},\n      {\"term_id\": \"R-HSA-5357801\", \"supporting_discovery_ids\": [0, 6, 13]}\n    ],\n    \"complexes\": [],\n    \"partners\": [\n      \"AKT1\",\n      \"SIRT1\",\n      \"PML\",\n      \"AR\",\n      \"CREBBP\",\n      \"DYRK1A\",\n      \"NLK\",\n      \"SCP4\"\n    ],\n    \"other_free_text\": []\n  }\n}\n```\n\n--- ROUND 2 (R2/R3, augmented corpus) ---\n\n```json\n{\n  \"mechanistic_narrative\": \"FOXO1 is a forkhead-family transcription factor that serves as a central integrator of insulin/PI3K/Akt signaling with metabolic gene regulation, cell-cycle control, apoptosis, autophagy, and inflammatory programs across diverse cell types. Akt directly phosphorylates FOXO1 at Thr-24, Ser-256, and Ser-319, triggering nuclear exclusion, Skp2-mediated ubiquitination, and proteasomal degradation; additional kinases (CK1, DYRK1A, NLK, PAK1) and the phosphatase SCP4 fine-tune its subcellular localization, while CBP-mediated acetylation attenuates DNA binding and SIRT1/SIRT2-mediated deacetylation restores transcriptional activity or redirects FOXO1 to a cytoplasmic Atg7-dependent autophagy pathway, and PRMT1-mediated arginine methylation antagonizes Akt phosphorylation [PMID:10358014, PMID:13679577, PMID:15668399, PMID:14710188, PMID:15220471, PMID:16076959, PMID:18951090, PMID:20543840, PMID:28851713]. In the nucleus, FOXO1 binds insulin-response elements to activate gluconeogenic genes (G6Pase, PEPCK), pro-apoptotic targets (FasL, PUMA, GADD45α), and immune-regulatory genes (TLR4, CCR7, IL-9), while repressing D-type cyclins, MYC, and eNOS to enforce cell-cycle arrest, endothelial quiescence, and metabolic restraint [PMID:11696581, PMID:12150827, PMID:26735015, PMID:21045807, PMID:25786691, PMID:16100571]. Foxo1-null mice die at ~E11 from angiogenic defects, and tissue-specific deletions in liver, endothelium, macrophages, intestinal epithelium, and immune cells demonstrate non-redundant roles in gluconeogenesis, vascular remodeling, inflammatory polarization, goblet cell autophagy, and T-cell differentiation [PMID:15184386, PMID:22147007, PMID:35700043, PMID:34287641, PMID:29339772].\",\n  \"teleology\": [\n    {\n      \"year\": 1999,\n      \"claim\": \"Identification of Akt as the direct kinase for FOXO1 at three conserved sites established the core mechanism by which insulin/PI3K signaling inactivates FOXO1 transcriptional activity and triggers apoptosis when this regulation is lost.\",\n      \"evidence\": \"In vitro kinase assay with purified Akt, alanine-substitution mutagenesis, reporter and apoptosis assays in mammalian cells\",\n      \"pmids\": [\"10358014\", \"10358075\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Structural basis of Akt–FOXO1 interaction not resolved\", \"In vivo phosphorylation kinetics not determined\"]\n    },\n    {\n      \"year\": 1999,\n      \"claim\": \"Demonstration that FOXO1 binds insulin-response sequences and activates IRS-containing promoters defined it as a transcription factor directly mediating insulin-responsive gene regulation.\",\n      \"evidence\": \"In vitro DNA-binding assay and luciferase reporter in mammalian cells\",\n      \"pmids\": [\"10358014\", \"10358075\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Genome-wide binding profile not yet mapped\", \"Endogenous target genes not identified at this stage\"]\n    },\n    {\n      \"year\": 2001,\n      \"claim\": \"Gain- and loss-of-function experiments in hepatocytes established FOXO1 as the mediator of insulin regulation of gluconeogenic genes (G6Pase, PEPCK), connecting FOXO1 phosphorylation to whole-body glucose homeostasis.\",\n      \"evidence\": \"Adenoviral FOXO1 transduction and dominant-negative mutant in primary hepatocytes and kidney epithelial cells\",\n      \"pmids\": [\"11696581\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Liver-specific in vivo deletion not yet performed\", \"Relative contribution of FOXO1 versus other FOXO paralogs in hepatic gluconeogenesis unknown\"]\n    },\n    {\n      \"year\": 2001,\n      \"claim\": \"Discovery that DYRK1A phosphorylates FOXO1 at Ser-329 to promote nuclear exclusion revealed a PI3K-independent input controlling FOXO1 activity.\",\n      \"evidence\": \"Purified kinase from skeletal muscle, co-IP, nuclear co-localization, site-directed mutagenesis\",\n      \"pmids\": [\"11311120\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Physiological contexts requiring DYRK1A–FOXO1 regulation not defined\", \"In vivo significance not validated with DYRK1A knockout\"]\n    },\n    {\n      \"year\": 2002,\n      \"claim\": \"ChIP-validated occupancy of cyclin D promoters showed FOXO1 enforces cell-cycle arrest through transcriptional repression of D-type cyclins rather than solely through gene activation, broadening the mechanistic repertoire.\",\n      \"evidence\": \"Transcriptional profiling, ChIP, cell-cycle and transformation assays\",\n      \"pmids\": [\"12150827\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Mechanism of transcriptional repression (corepressor identity) not determined\"]\n    },\n    {\n      \"year\": 2003,\n      \"claim\": \"Reconstitution of phosphorylation-dependent ubiquitination and proteasomal degradation of FOXO1 established a two-step inactivation model (nuclear exclusion then degradation) downstream of insulin/Akt, with Skp2 identified as the E3 ligase.\",\n      \"evidence\": \"In vitro and in vivo ubiquitination assays with proteasome inhibitors and phospho-site mutants; Skp2 co-IP and in vitro ubiquitination\",\n      \"pmids\": [\"13679577\", \"15668399\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether additional E3 ligases target FOXO1 in specific tissues not resolved\", \"Deubiquitinases for FOXO1 not identified\"]\n    },\n    {\n      \"year\": 2004,\n      \"claim\": \"CK1-mediated phosphorylation at Ser-322/325 was shown to cooperate with Akt phosphorylation to drive nuclear exclusion, revealing multisite phosphorylation as a combinatorial regulatory code.\",\n      \"evidence\": \"CK1 inhibitor D4476, phospho-specific analysis, nuclear/cytoplasmic fractionation in hepatoma cells\",\n      \"pmids\": [\"14710188\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"CK1 isoform specificity not fully determined\", \"Interplay with DYRK1A and NLK phosphorylation not tested\"]\n    },\n    {\n      \"year\": 2004,\n      \"claim\": \"Foxo1-null embryonic lethality at ~E11 with angiogenic defects established FOXO1 as essential for vascular development and endothelial VEGF responses.\",\n      \"evidence\": \"Constitutive Foxo1 knockout mice, ES-cell-derived endothelial differentiation assay\",\n      \"pmids\": [\"15184386\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Downstream transcriptional targets mediating the vascular phenotype not fully defined at this stage\"]\n    },\n    {\n      \"year\": 2004,\n      \"claim\": \"Demonstration that SIRT1 deacetylates FOXO1 at CBP-acetylated lysines and is recruited to FOXO1 target promoters established the acetylation/deacetylation switch as a major regulatory axis modulating FOXO1 transcriptional output.\",\n      \"evidence\": \"In vitro deacetylation with purified SIRT1, ChIP on MnSOD/p27 promoters, catalytic-dead mutant control\",\n      \"pmids\": [\"15220471\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Context-dependent selection of FOXO1 targets by acetylation state not systematically mapped\"]\n    },\n    {\n      \"year\": 2005,\n      \"claim\": \"Biochemical reconstitution showed CBP acetylation at Lys-242/245/262 attenuates FOXO1 DNA binding by charge neutralization and sensitizes FOXO1 to Akt phosphorylation, revealing bidirectional crosstalk between acetylation and phosphorylation.\",\n      \"evidence\": \"In vitro DNA binding and kinase assays with acetylation-mimicking and acetylation-deficient mutants\",\n      \"pmids\": [\"16076959\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"In vivo stoichiometry of acetylated versus unacetylated FOXO1 unknown\"]\n    },\n    {\n      \"year\": 2005,\n      \"claim\": \"Discovery that FOXO1 forms a complex with PML and SIRT1 in pancreatic beta cells to activate NeuroD/MafA transcription under oxidative stress revealed a tissue-specific protective function coupling acetylation to PML body localization.\",\n      \"evidence\": \"Co-IP of FoxO1–PML–SIRT1 complex, acetylation mutants, transgenic mice with beta-cell phenotyping\",\n      \"pmids\": [\"16154098\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether PML–SIRT1–FOXO1 complex operates in non-beta-cell contexts not tested\"]\n    },\n    {\n      \"year\": 2005,\n      \"claim\": \"Beta-catenin was identified as a direct FOXO co-activator under oxidative stress, conserved from C. elegans to mammals, linking Wnt pathway components to FOXO-dependent stress responses.\",\n      \"evidence\": \"Co-IP, reporter assay, C. elegans genetic epistasis (BAR-1/DAF-16)\",\n      \"pmids\": [\"15905404\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Competition between TCF/LEF and FOXO for beta-catenin binding not quantitatively resolved\"]\n    },\n    {\n      \"year\": 2006,\n      \"claim\": \"Liver-specific constitutively active FOXO1 transgenics demonstrated that FOXO1 globally coordinates hepatic metabolism by activating gluconeogenesis while suppressing glycolytic, lipogenic, and pentose phosphate pathway genes.\",\n      \"evidence\": \"Transgenic mice with liver-specific active FoxO1, gene array, adenoviral hepatocyte transduction, metabolic phenotyping\",\n      \"pmids\": [\"16492665\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Direct versus indirect transcriptional targets within suppressed metabolic programs not distinguished\"]\n    },\n    {\n      \"year\": 2008,\n      \"claim\": \"PRMT1-mediated methylation at Arg-248/250 was shown to directly block Akt phosphorylation at Ser-253, establishing arginine methylation as a third post-translational code that antagonizes the Akt-ubiquitination axis.\",\n      \"evidence\": \"In vitro methylation and kinase assays, PRMT1 siRNA, polyubiquitination assay, apoptosis assay\",\n      \"pmids\": [\"18951090\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Physiological stimuli triggering PRMT1-dependent FOXO1 methylation not defined\", \"Demethylase not identified\"]\n    },\n    {\n      \"year\": 2010,\n      \"claim\": \"ChIP-Seq in macrophages revealed FOXO1 occupancy at enhancer-like elements within TLR4 and downstream innate immune genes, with TLR4 activation feeding back through Akt to inactivate FOXO1—establishing a self-limiting inflammatory circuit.\",\n      \"evidence\": \"ChIP-Seq, FoxO1 knockout macrophages, reporter assays, cytokine measurements\",\n      \"pmids\": [\"21045807\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Precise chromatin remodeling events at FOXO1-bound enhancers not characterized\"]\n    },\n    {\n      \"year\": 2010,\n      \"claim\": \"Cytoplasmic acetylated FOXO1 was shown to bind Atg7 and induce autophagy independently of transcription, revealing a non-nuclear tumor-suppressive function mediated by SIRT2 dissociation.\",\n      \"evidence\": \"Co-IP (FoxO1–Atg7), transcription-deficient mutants, autophagy assays, SIRT2 interaction studies, xenograft tumor model\",\n      \"pmids\": [\"20543840\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Structural basis of acetylated FOXO1–Atg7 interaction unknown\", \"Relative contribution of transcriptional versus cytoplasmic autophagy pathways in different tissues not quantified\"]\n    },\n    {\n      \"year\": 2011,\n      \"claim\": \"Liver-specific Foxo1 deletion confirmed its non-redundant role in gluconeogenesis in vivo, while combined Foxo1/Foxo3 deletion additionally derepressed lipogenesis, genetically separating paralog-specific metabolic functions.\",\n      \"evidence\": \"Cre/LoxP conditional single and combined knockouts, metabolic phenotyping, hepatic transcript analysis\",\n      \"pmids\": [\"22147007\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Mechanism by which Foxo3 specifically suppresses lipogenic genes independently of Foxo1 not fully delineated\"]\n    },\n    {\n      \"year\": 2016,\n      \"claim\": \"Endothelial-specific genetic models demonstrated FOXO1 enforces vascular quiescence by repressing MYC, restraining glycolysis and proliferation; MYC re-expression rescued the FOXO1-overexpression phenotype, defining a FOXO1–MYC metabolic antagonism in angiogenesis.\",\n      \"evidence\": \"Endothelial-specific knockout and overexpression mice, metabolic flux assays, MYC epistasis rescue\",\n      \"pmids\": [\"26735015\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether FOXO1 directly represses MYC transcription or acts post-transcriptionally not resolved\"]\n    },\n    {\n      \"year\": 2017,\n      \"claim\": \"Identification of SCP4 as the phosphatase that directly dephosphorylates FOXO1 to retain it in the nucleus provided the missing counter-regulatory enzyme to Akt; SCP4-null mice exhibit neonatal hypoglycemia, validating the axis in vivo.\",\n      \"evidence\": \"In vitro dephosphorylation assay, SCP4 knockout mice, hepatic glucose production assays\",\n      \"pmids\": [\"28851713\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Regulation of SCP4 itself under fasting/feeding transitions not characterized\"]\n    },\n    {\n      \"year\": 2020,\n      \"claim\": \"FOXO1 was shown to directly bind the KLF5 promoter in cardiomyocytes, initiating a KLF5→NOX4→ROS→ceramide cascade underlying diabetic cardiomyopathy; cardiomyocyte-specific FOXO1 deletion was protective.\",\n      \"evidence\": \"ChIP (FOXO1 on KLF5 promoter), cardiomyocyte-specific knockout, AAV rescue, oxidative stress measurements\",\n      \"pmids\": [\"33539225\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether FOXO1-KLF5 axis operates in non-diabetic cardiac stress unknown\"]\n    },\n    {\n      \"year\": 2022,\n      \"claim\": \"Myeloid-specific FOXO1 deletion revealed its role in promoting M1 macrophage polarization by opposing Stat6 signaling, connecting FOXO1 to inflammatory hepatic pathology in NASH.\",\n      \"evidence\": \"Myeloid-specific Foxo1 conditional knockout (LysM-Cre), M1/M2 markers, Stat6 pathway analysis, NASH/HFD models\",\n      \"pmids\": [\"35700043\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Molecular mechanism of FOXO1–Stat6 antagonism (direct binding vs. transcriptional) not defined\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"Key unresolved questions include how tissue-specific cofactor assemblies select distinct FOXO1 transcriptional programs, the structural basis for the acetylated-FOXO1–Atg7 autophagy complex, the identity of FOXO1 demethylases and additional deubiquitinases, and how the multiple kinase inputs (Akt, CK1, DYRK1A, NLK, PAK1, cGKI, mTORC2/HDAC) are integrated in real time at the single-cell level.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Low\",\n      \"gaps\": [\"No high-resolution structure of full-length FOXO1 with cofactors\", \"Integrative quantitative modeling of multisite post-translational modification crosstalk lacking\", \"Tissue-specific FOXO1 interactome not systematically mapped\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0140110\", \"supporting_discovery_ids\": [1, 2, 4, 9, 11, 14, 18, 22, 29, 31, 35, 38]},\n      {\"term_id\": \"GO:0003677\", \"supporting_discovery_ids\": [1, 4, 12, 22, 29, 35, 38, 43]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005634\", \"supporting_discovery_ids\": [0, 3, 7, 9, 11, 12, 20, 25, 33]},\n      {\"term_id\": \"GO:0005829\", \"supporting_discovery_ids\": [5, 7, 15, 24]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"GO:0162582\", \"supporting_discovery_ids\": [0, 5, 7, 13, 20, 27, 33]},\n      {\"term_id\": \"R-HSA-74160\", \"supporting_discovery_ids\": [1, 2, 4, 18, 22, 29, 31, 38]},\n      {\"term_id\": \"R-HSA-1430728\", \"supporting_discovery_ids\": [2, 18, 26, 36, 43]},\n      {\"term_id\": \"R-HSA-162582\", \"supporting_discovery_ids\": [0, 5, 7, 13, 20, 27, 33]},\n      {\"term_id\": \"R-HSA-5357801\", \"supporting_discovery_ids\": [0, 6, 13, 20, 25]},\n      {\"term_id\": \"R-HSA-9612973\", \"supporting_discovery_ids\": [24, 39, 41]},\n      {\"term_id\": \"R-HSA-1640170\", \"supporting_discovery_ids\": [4]},\n      {\"term_id\": \"R-HSA-168256\", \"supporting_discovery_ids\": [22, 29, 35, 42]},\n      {\"term_id\": \"R-HSA-392499\", \"supporting_discovery_ids\": [5, 13, 20]}\n    ],\n    \"complexes\": [\n      \"PML-SIRT1-FOXO1\",\n      \"FOXO1-Atg7\",\n      \"FOXO1-beta-catenin\"\n    ],\n    \"partners\": [\n      \"AKT1\",\n      \"SIRT1\",\n      \"SIRT2\",\n      \"SKP2\",\n      \"PRMT1\",\n      \"CTNNB1\",\n      \"ATG7\",\n      \"PML\"\n    ],\n    \"other_free_text\": []\n  }\n}\n```"}