{"gene":"FLOT1","run_date":"2026-04-28T17:46:03","timeline":{"discoveries":[{"year":1997,"finding":"Flotillin-1 was identified and molecularly cloned as a resident integral membrane protein component of caveolae, localizing to the Triton-insoluble, buoyant membrane fraction (lipid rafts) in brain and other tissues, and was shown to define a novel family of caveolae-associated integral membrane proteins together with epidermal surface antigen (flotillin-2).","method":"Molecular cloning, detergent-resistant membrane fractionation, multiple independent biochemical methods confirming caveolar localization","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 2 — multiple orthogonal biochemical methods, foundational characterization paper, highly cited","pmids":["9153235"],"is_preprint":false},{"year":2000,"finding":"Flotillin-1 was identified as forming a ternary complex with CAP (c-Cbl-associated protein) and Cbl, directing the localization of the CAP-Cbl complex to a lipid raft subdomain of the plasma membrane upon insulin stimulation; this pathway was shown to be essential for insulin-stimulated glucose transport independent of PI3K signaling.","method":"Yeast two-hybrid screen, co-immunoprecipitation, dominant-negative overexpression in 3T3-L1 adipocytes, glucose transport assay","journal":"Nature","confidence":"High","confidence_rationale":"Tier 2 — reciprocal Co-IP, functional rescue, defined cellular phenotype; highly cited foundational paper","pmids":["11001060"],"is_preprint":false},{"year":2001,"finding":"Flotillin-1 was identified as a major integral protein of human erythrocyte lipid rafts, forming independently organized high-order oligomers that act as separate scaffolding components at the cytoplasmic face of erythrocyte lipid rafts.","method":"Lipid raft isolation from human erythrocytes, protein identification, sucrose gradient fractionation, oligomer characterization","journal":"Blood","confidence":"High","confidence_rationale":"Tier 2 — direct biochemical isolation and characterization with multiple methods; highly cited","pmids":["11159550"],"is_preprint":false},{"year":2005,"finding":"Flotillin-1 defines a clathrin-independent endocytic pathway in mammalian cells: it resides in punctate plasma membrane structures distinct from clathrin-coated pits and caveolin-1-positive caveolae, accumulates GPI-linked proteins and cholera toxin B subunit in endocytic intermediates, and siRNA knockdown inhibits clathrin-independent uptake of cholera toxin and GPI-linked protein endocytosis.","method":"Ferro-fluid-based endosome purification, total internal reflection microscopy, immuno-electron microscopy, siRNA knockdown, endocytosis assays","journal":"Nature cell biology","confidence":"High","confidence_rationale":"Tier 2 — multiple orthogonal methods including live imaging, immuno-EM, and functional siRNA knockdown; highly cited","pmids":["16341206"],"is_preprint":false},{"year":2007,"finding":"Coassembly of flotillin-1 and flotillin-2 is sufficient to generate de novo membrane microdomains distinct from caveolin-1-positive caveolae; this coassembly induces membrane curvature, formation of plasma-membrane invaginations, and accumulation of intracellular vesicles, establishing flotillin-1 as a defining structural component of a clathrin-independent endocytic machinery.","method":"Overexpression of flotillin constructs, live-cell imaging, electron microscopy, membrane curvature and vesicle budding assays","journal":"Current biology : CB","confidence":"High","confidence_rationale":"Tier 2 — reconstitution-like overexpression system with multiple orthogonal readouts including EM; highly cited","pmids":["17600709"],"is_preprint":false},{"year":2011,"finding":"Silencing FLOT1 in breast cancer cells inhibited proliferation and tumorigenicity both in vitro and in vivo, mechanistically associated with suppression of Akt activity, enhanced transcriptional activity of FOXO3a, upregulation of p21(Cip1) and p27(Kip1), and downregulation of cyclin D1.","method":"siRNA knockdown, Western blotting, luciferase reporter assay, in vitro proliferation assays, in vivo xenograft","journal":"Clinical cancer research","confidence":"Medium","confidence_rationale":"Tier 2 — clean KD with multiple downstream pathway readouts in vitro and in vivo; single lab","pmids":["21447726"],"is_preprint":false},{"year":2017,"finding":"FLOT1 interacts with syndecan-1 (but not caveolin-1) in liver cells via the transmembrane/cytoplasmic region of syndecan-1 and the N-terminal hydrophobic domain of FLOT1; C-TRL binding to syndecan-1 enhances this association and the two proteins traffic together into lysosomes; FLOT1 knockdown substantially inhibits syndecan-1 endocytosis; adenoviral restoration of wild-type but not N-terminal hydrophobic domain-deleted FLOT1 in diabetic mice normalized plasma triglycerides.","method":"Co-immunoprecipitation, domain deletion mutants, siRNA knockdown, adenoviral overexpression in mouse model, plasma triglyceride assay","journal":"Arteriosclerosis, thrombosis, and vascular biology","confidence":"High","confidence_rationale":"Tier 2 — reciprocal Co-IP, domain-mapping mutagenesis, in vivo rescue with domain mutant; multiple orthogonal methods","pmids":["29162604"],"is_preprint":false},{"year":2019,"finding":"FLOT1 promotes lung adenocarcinoma cell growth, invasion, and migration, inhibits apoptosis, induces epithelial-mesenchymal transition, and modulates the cell cycle by activating the Erk/Akt signaling pathway.","method":"Lentiviral knockdown and overexpression, cell growth/invasion/migration assays, apoptosis assay, EMT marker analysis, Erk/Akt pathway analysis","journal":"Thoracic cancer","confidence":"Medium","confidence_rationale":"Tier 3 — KD/OE with phenotype and pathway markers but no direct binding partner or epistasis; single lab","pmids":["30838797"],"is_preprint":false},{"year":2023,"finding":"FLOT1 promotes gastric cancer progression and metastasis by physically interacting with BCAR1, regulating BCAR1 phosphorylation and translocation; BCAR1 knockdown blocks FLOT1-induced proliferation/migration/invasion; re-expression of wild-type but not BCAR1(Y410F) partially restores FLOT1-knockdown phenotypes, and this restoration is blocked by ERK inhibitor, placing FLOT1 upstream of BCAR1 phosphorylation and ERK signaling.","method":"Co-immunoprecipitation, overexpression/knockdown, BCAR1 phosphorylation-site mutant (Y410F), ERK inhibitor, proliferation/migration/invasion assays","journal":"International journal of biological sciences","confidence":"Medium","confidence_rationale":"Tier 2 — Co-IP plus phospho-site mutant rescue and pharmacological inhibition; single lab","pmids":["37928269"],"is_preprint":false},{"year":2023,"finding":"After irradiation, FLOT1 forms part of a SDC1-TGM2-FLOT1-BHMT complex that mediates autophagosome-lysosome fusion in GBM cells: SDC1 carries TGM2 from the cell membrane into the cytoplasm and transports it to lysosomes by binding to FLOT1, then TGM2 recognizes BHMT on autophagosomes to coordinate their encounter with lysosomes, maintaining autophagic flux and enhancing radioresistance.","method":"Co-immunoprecipitation, immunofluorescence, mRFP-GFP-LC3 assay, transmission electron microscopy, flow cytometry, Western blotting, qPCR","journal":"Theranostics","confidence":"Medium","confidence_rationale":"Tier 2 — Co-IP defining complex, functional assays with multiple readouts; single lab","pmids":["37441590"],"is_preprint":false},{"year":2023,"finding":"EIF4A3 physically interacts with FLOT1 in lung adenocarcinoma cells and positively regulates FLOT1 protein expression; FLOT1 knockdown reverses the increase in cell proliferation and migration caused by EIF4A3 overexpression and rescues EIF4A3-driven activation of the PI3K-AKT-ERK1/2-P70S6K signaling pathway and PI3K class III-mediated autophagy.","method":"Mass spectrometry (co-IP pull-down), transcriptome sequencing, siRNA knockdown, overexpression, cell proliferation/migration assays, pathway analysis by Western blot","journal":"Molecular cancer research : MCR","confidence":"Medium","confidence_rationale":"Tier 2 — MS-confirmed interaction, epistasis by rescue experiment; single lab","pmids":["37011005"],"is_preprint":false},{"year":2023,"finding":"FLOT1 knockdown in AML cells triggers both apoptosis and pyroptosis, inhibits tumor engraftment in vivo, while FLOT1 overexpression promotes AML cell growth and apoptosis resistance, demonstrating a role for FLOT1 in regulating cell death pathways in hematological malignancy.","method":"siRNA knockdown, overexpression, flow cytometry, in vivo xenograft engraftment assay, Western blotting","journal":"Annals of hematology","confidence":"Medium","confidence_rationale":"Tier 2 — KD/OE with defined cell death phenotypes in vitro and in vivo; single lab","pmids":["36697954"],"is_preprint":false},{"year":2025,"finding":"SMARCC1 activates FLOT1 transcription by binding to its promoter; FLOT1 promotes M2 macrophage polarization, increases PD-L1 expression, and reduces ferroptosis in macrophages by restoring GSH:GSSG ratio and preventing lipid peroxidation; FLOT1 overexpression rescues the inhibitory effects of SMARCC1 knockdown on M2 macrophage infiltration and ferroptosis suppression.","method":"ChIP assay (SMARCC1 binding to FLOT1 promoter), siRNA knockdown, overexpression, GSH/GSSG ratio measurement, lipid peroxidation assay, transmission electron microscopy of mitochondria, co-culture assays, xenograft models","journal":"Journal of molecular medicine (Berlin, Germany)","confidence":"Medium","confidence_rationale":"Tier 2 — ChIP-confirmed transcriptional regulation, multiple downstream functional assays; single lab","pmids":["40108025"],"is_preprint":false},{"year":2026,"finding":"In Alzheimer's disease models, FLOT1 interacts with the transcription factor FOSL2, which upregulates EphA2 expression, leading to activation of the p38/MAPK signaling pathway and pro-inflammatory polarization of microglia; silencing FLOT1 in APP/PS1 mice reduced neuroinflammatory markers, prevented pro-inflammatory microglial polarization, and improved spatial memory.","method":"Co-immunoprecipitation (FLOT1-FOSL2 interaction), ChIP assay (FOSL2 on EphA2 promoter), dual-luciferase assay, qPCR, Western blotting, IHC, Morris water maze in APP/PS1 mice","journal":"Neuropharmacology","confidence":"Medium","confidence_rationale":"Tier 2 — Co-IP, ChIP, luciferase, and in vivo behavioral rescue; single lab","pmids":["41548752"],"is_preprint":false},{"year":2026,"finding":"Flotillin-1 (Flot1) localizes to circular dorsal ruffles (CDRs) in podocytes; Flot1 depletion reduces macropinosome formation and impairs growth-factor-stimulated mTORC1 activation, demonstrating that Flot1 participates in CDR-derived macropinosome formation and macropinosome-dependent nutrient delivery to lysosomes for mTORC1 activation.","method":"Imaging analysis (Flot1 localization at CDRs), Flot1 knockout cells, macropinosome formation assay, mTORC1 activation biochemical assay, growth assay, immunostaining","journal":"Cell structure and function","confidence":"Medium","confidence_rationale":"Tier 2 — KO with direct localization data and mechanistic mTORC1 signaling readout; single lab","pmids":["41500583"],"is_preprint":false},{"year":2020,"finding":"FLOT1 mRNA is regulated by N6-methyladenosine (m6A) modification in ovarian cancer cells; the level of m6A modification of FLOT1 mRNA is significantly elevated in OC cells compared with normal cells, leading to increased FLOT1 mRNA expression; application of the methylation inhibitor 3-deazaadenosine decreased FLOT1 mRNA expression and suppressed tumor formation in a xenograft model.","method":"m6A modification analysis, methylation inhibitor treatment, qRT-PCR, xenograft mouse model","journal":"Cell biology international","confidence":"Low","confidence_rationale":"Tier 3 — m6A association shown but writer/reader/eraser not specifically identified; single lab","pmids":["40066501"],"is_preprint":false}],"current_model":"FLOT1 is a lipid raft scaffold protein that drives clathrin-independent endocytosis by coassembling with flotillin-2 to generate membrane microdomains that bud into the cell; it mediates syndecan-1-dependent lipoprotein disposal via its N-terminal hydrophobic domain, forms a ternary complex with CAP and Cbl to direct insulin-stimulated glucose transport, promotes cancer cell proliferation and survival through Akt/FOXO3a and ERK signaling (including via BCAR1 interaction), participates in autophagosome-lysosome fusion as part of an SDC1-TGM2-FLOT1-BHMT complex, regulates mTORC1 activation through macropinosome formation in podocytes, and in the brain interacts with FOSL2 to drive EphA2/p38-MAPK-dependent pro-inflammatory microglial polarization relevant to Alzheimer's disease."},"narrative":{"teleology":[{"year":1997,"claim":"Establishing that FLOT1 is an integral membrane protein of caveolae/lipid rafts defined a new structural component of detergent-resistant membrane domains and founded the flotillin protein family.","evidence":"Molecular cloning and detergent-resistant membrane fractionation in brain and other tissues","pmids":["9153235"],"confidence":"High","gaps":["Whether FLOT1 was functionally distinct from caveolins was unknown","No endocytic or signaling role yet assigned"]},{"year":2000,"claim":"Discovery that FLOT1 scaffolds a CAP–Cbl ternary complex at lipid rafts to enable insulin-stimulated glucose transport established FLOT1's first signaling function and linked it to metabolic regulation.","evidence":"Yeast two-hybrid, reciprocal co-immunoprecipitation, dominant-negative overexpression in 3T3-L1 adipocytes with glucose transport assay","pmids":["11001060"],"confidence":"High","gaps":["Structural basis of the FLOT1–CAP interaction not determined","Whether this pathway operates in vivo in whole-animal insulin signaling was untested"]},{"year":2001,"claim":"Demonstrating that FLOT1 forms high-order oligomers as an independent scaffolding platform in erythrocyte lipid rafts established that its structural role extends beyond caveolin-containing membranes.","evidence":"Lipid raft isolation from human erythrocytes, sucrose gradient fractionation, oligomer characterization","pmids":["11159550"],"confidence":"High","gaps":["Oligomerization interfaces and stoichiometry undefined","Functional consequence of oligomerization for membrane trafficking not yet tested"]},{"year":2005,"claim":"Identification of FLOT1-positive puncta as a clathrin- and caveolin-independent endocytic pathway resolved the question of whether lipid raft endocytosis requires a distinct coat-like machinery.","evidence":"TIRF microscopy, immuno-EM, ferro-fluid endosome purification, siRNA knockdown inhibiting GPI-linked protein and cholera toxin uptake","pmids":["16341206"],"confidence":"High","gaps":["Cargo selectivity mechanism for flotillin-mediated endocytosis unknown","Dynamin dependence of the pathway not fully resolved"]},{"year":2007,"claim":"Showing that FLOT1–FLOT2 coassembly is sufficient to generate membrane curvature and vesicle budding established flotillins as minimal machinery for de novo microdomain formation and endocytosis.","evidence":"Overexpression of flotillin constructs with live-cell imaging and electron microscopy of induced invaginations and vesicles","pmids":["17600709"],"confidence":"High","gaps":["No in vitro reconstitution with purified proteins","Post-translational modifications controlling assembly not mapped"]},{"year":2011,"claim":"Linking FLOT1 to cancer cell proliferation through the Akt–FOXO3a axis expanded its role from membrane trafficking scaffold to a regulator of growth and survival signaling.","evidence":"siRNA knockdown in breast cancer cells, Western blot for Akt/FOXO3a targets, in vivo xenograft","pmids":["21447726"],"confidence":"Medium","gaps":["Direct mechanism by which FLOT1 activates Akt was not identified","Not confirmed in non-cancer primary cells"]},{"year":2017,"claim":"Mapping the FLOT1–syndecan-1 interaction to the N-terminal hydrophobic domain and showing that in vivo FLOT1 re-expression normalizes plasma triglycerides demonstrated a physiological role for flotillin-mediated endocytosis in lipoprotein metabolism.","evidence":"Co-immunoprecipitation with domain-deletion mutants, siRNA knockdown, adenoviral rescue with WT vs. ΔN-FLOT1 in diabetic mice","pmids":["29162604"],"confidence":"High","gaps":["Whether FLOT1 directly contacts lipoprotein particles or acts solely via SDC1 remains unclear","Structural details of the N-terminal hydrophobic domain interaction undefined"]},{"year":2023,"claim":"Multiple 2023 studies extended FLOT1's signaling roles: FLOT1 phosphorylates BCAR1 to drive ERK-dependent gastric cancer progression, participates in an SDC1–TGM2–FLOT1–BHMT complex for autophagosome–lysosome fusion in glioblastoma, acts downstream of EIF4A3 to relay PI3K–AKT signaling in lung adenocarcinoma, and regulates apoptosis/pyroptosis in AML cells.","evidence":"Co-IP with phospho-site mutant rescue and ERK inhibitor (gastric cancer); mRFP-GFP-LC3 flux assay and TEM (GBM autophagy); MS-confirmed interaction and epistasis rescue (LUAD); KD/OE with xenograft (AML)","pmids":["37928269","37441590","37011005","36697954"],"confidence":"Medium","gaps":["Kinase that phosphorylates BCAR1 downstream of FLOT1 not identified","Stoichiometry and assembly order of the SDC1–TGM2–FLOT1–BHMT complex not established","Most cancer studies from single laboratories and lack independent replication"]},{"year":2025,"claim":"Demonstrating that FLOT1 promotes M2 macrophage polarization, PD-L1 upregulation, and ferroptosis resistance by maintaining GSH:GSSG balance extended FLOT1 function to immune-cell regulation and redox homeostasis.","evidence":"ChIP for SMARCC1 on FLOT1 promoter, GSH/GSSG ratio measurement, lipid peroxidation assay, co-culture and xenograft models","pmids":["40108025"],"confidence":"Medium","gaps":["Direct molecular mechanism linking FLOT1 to GSH metabolism is unknown","Single laboratory; not replicated"]},{"year":2026,"claim":"Two 2026 studies revealed tissue-specific FLOT1 functions: interaction with FOSL2 driving EphA2/p38-MAPK–dependent pro-inflammatory microglial polarization in Alzheimer's disease, and localization to circular dorsal ruffles in podocytes to support macropinosome-dependent mTORC1 activation.","evidence":"Co-IP and ChIP for FOSL2–EphA2, Morris water maze in APP/PS1 mice (AD); Flot1 KO podocytes with macropinosome and mTORC1 assays","pmids":["41548752","41500583"],"confidence":"Medium","gaps":["How FLOT1 recruits or stabilizes FOSL2 mechanistically is undefined","Whether the macropinosome–mTORC1 axis is specific to podocytes or general has not been tested","Both findings from single laboratories"]},{"year":null,"claim":"A high-resolution structural model of the FLOT1–FLOT2 oligomeric scaffold, the identity of cargo-selectivity determinants, and the direct mechanism linking FLOT1 to Akt and mTOR activation remain unresolved.","evidence":"","pmids":[],"confidence":"High","gaps":["No crystal or cryo-EM structure of FLOT1 or the FLOT1–FLOT2 complex","Cargo selectivity rules for flotillin-mediated endocytosis not determined","Direct kinase/effector connecting FLOT1 scaffolding to Akt phosphorylation unknown"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0008289","term_label":"lipid binding","supporting_discovery_ids":[0,2,3]},{"term_id":"GO:0060090","term_label":"molecular adaptor activity","supporting_discovery_ids":[1,6,9]},{"term_id":"GO:0005198","term_label":"structural molecule activity","supporting_discovery_ids":[2,4]}],"localization":[{"term_id":"GO:0005886","term_label":"plasma membrane","supporting_discovery_ids":[0,1,2,3,4,14]},{"term_id":"GO:0031410","term_label":"cytoplasmic vesicle","supporting_discovery_ids":[3,4,6,14]},{"term_id":"GO:0005764","term_label":"lysosome","supporting_discovery_ids":[6,9]},{"term_id":"GO:0005829","term_label":"cytosol","supporting_discovery_ids":[9]}],"pathway":[{"term_id":"R-HSA-5653656","term_label":"Vesicle-mediated transport","supporting_discovery_ids":[3,4,6,14]},{"term_id":"R-HSA-162582","term_label":"Signal Transduction","supporting_discovery_ids":[1,5,7,8,13]},{"term_id":"R-HSA-9612973","term_label":"Autophagy","supporting_discovery_ids":[9,10]},{"term_id":"R-HSA-168256","term_label":"Immune System","supporting_discovery_ids":[12,13]},{"term_id":"R-HSA-5357801","term_label":"Programmed Cell Death","supporting_discovery_ids":[11,12]}],"complexes":["FLOT1-FLOT2 oligomeric scaffold","CAP-Cbl-FLOT1 ternary complex","SDC1-TGM2-FLOT1-BHMT complex"],"partners":["FLOT2","SORBS1","CBL","SDC1","TGM2","BCAR1","FOSL2","EIF4A3"],"other_free_text":[]},"mechanistic_narrative":"FLOT1 is a lipid-raft scaffold protein that organizes membrane microdomains and drives clathrin-independent endocytosis, linking membrane trafficking to diverse signaling and metabolic outcomes. FLOT1 homo- and hetero-oligomerizes with flotillin-2 to induce membrane curvature and vesicle budding independently of clathrin and caveolin, mediating uptake of GPI-anchored proteins, cholera toxin, and syndecan-1-dependent triglyceride-rich lipoproteins [PMID:9153235, PMID:16341206, PMID:17600709, PMID:29162604]. Through its scaffolding function, FLOT1 nucleates signaling complexes—including a CAP–Cbl ternary complex required for insulin-stimulated glucose uptake and a BCAR1-dependent ERK cascade in cancer cells—and participates in autophagosome–lysosome fusion as part of an SDC1–TGM2–FLOT1–BHMT complex and in macropinosome-dependent mTORC1 activation in podocytes [PMID:11001060, PMID:37928269, PMID:37441590, PMID:41500583]. FLOT1 also modulates immune cell phenotypes, promoting M2 macrophage polarization with ferroptosis resistance and driving pro-inflammatory microglial polarization via a FOSL2–EphA2–p38/MAPK axis relevant to Alzheimer's disease pathology [PMID:40108025, PMID:41548752]."},"prefetch_data":{"uniprot":{"accession":"O75955","full_name":"Flotillin-1","aliases":[],"length_aa":427,"mass_kda":47.4,"function":"May act as a scaffolding protein within caveolar membranes, functionally participating in formation of caveolae or caveolae-like vesicles","subcellular_location":"Cell membrane; Endosome; Membrane, caveola; Melanosome; Membrane raft","url":"https://www.uniprot.org/uniprotkb/O75955/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/FLOT1","classification":"Not Classified","n_dependent_lines":71,"n_total_lines":1208,"dependency_fraction":0.058774834437086095},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[{"gene":"CALD1","stoichiometry":0.2},{"gene":"RAB11A","stoichiometry":0.2},{"gene":"RANBP1","stoichiometry":0.2}],"url":"https://opencell.sf.czbiohub.org/search/FLOT1","total_profiled":1310},"omim":[{"mim_id":"616694","title":"ECM29 PROTEASOME ADAPTOR AND SCAFFOLD PROTEIN; ECPAS","url":"https://www.omim.org/entry/616694"},{"mim_id":"610195","title":"PTOV1 EXTENDED AT-HOOK-CONTAINING ADAPTOR PROTEIN; PTOV1","url":"https://www.omim.org/entry/610195"},{"mim_id":"608010","title":"NPC1-LIKE INTRACELLULAR CHOLESTEROL TRANSPORTER 1; NPC1L1","url":"https://www.omim.org/entry/608010"},{"mim_id":"606998","title":"FLOTILLIN 1; FLOT1","url":"https://www.omim.org/entry/606998"},{"mim_id":"602744","title":"GLYCERONEPHOSPHATE O-ACYLTRANSFERASE; GNPAT","url":"https://www.omim.org/entry/602744"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"Uncertain","locations":[{"location":"Golgi apparatus","reliability":"Uncertain"},{"location":"Vesicles","reliability":"Additional"},{"location":"Plasma membrane","reliability":"Additional"}],"tissue_specificity":"Low tissue specificity","tissue_distribution":"Detected in all","driving_tissues":[],"url":"https://www.proteinatlas.org/search/FLOT1"},"hgnc":{"alias_symbol":[],"prev_symbol":[]},"alphafold":{"accession":"O75955","domains":[{"cath_id":"-","chopping":"2-44","consensus_level":"medium","plddt":89.0653,"start":2,"end":44},{"cath_id":"3.30.479.30","chopping":"49-159","consensus_level":"high","plddt":92.5041,"start":49,"end":159}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/O75955","model_url":"https://alphafold.ebi.ac.uk/files/AF-O75955-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-O75955-F1-predicted_aligned_error_v6.png","plddt_mean":81.62},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=FLOT1","jax_strain_url":"https://www.jax.org/strain/search?query=FLOT1"},"sequence":{"accession":"O75955","fasta_url":"https://rest.uniprot.org/uniprotkb/O75955.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/O75955/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/O75955"}},"corpus_meta":[{"pmid":"22589463","id":"PMC_22589463","title":"A membrane microdomain-associated protein, Arabidopsis Flot1, is involved in a clathrin-independent endocytic pathway and is required for seedling development.","date":"2012","source":"The Plant cell","url":"https://pubmed.ncbi.nlm.nih.gov/22589463","citation_count":154,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"21447726","id":"PMC_21447726","title":"Knockdown of FLOT1 impairs cell proliferation and tumorigenicity in breast cancer through upregulation of FOXO3a.","date":"2011","source":"Clinical cancer research : an official journal of the American Association for Cancer Research","url":"https://pubmed.ncbi.nlm.nih.gov/21447726","citation_count":106,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"26002553","id":"PMC_26002553","title":"miRNA-target network reveals miR-124as a key miRNA contributing to clear cell renal cell carcinoma aggressive behaviour by targeting CAV1 and FLOT1.","date":"2015","source":"Oncotarget","url":"https://pubmed.ncbi.nlm.nih.gov/26002553","citation_count":72,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"25793370","id":"PMC_25793370","title":"MiR-506 is down-regulated in clear cell renal cell carcinoma and inhibits cell growth and metastasis via targeting FLOT1.","date":"2015","source":"PloS one","url":"https://pubmed.ncbi.nlm.nih.gov/25793370","citation_count":56,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"30771789","id":"PMC_30771789","title":"Integration of GWAS and brain eQTL identifies FLOT1 as a risk gene for major depressive disorder.","date":"2019","source":"Neuropsychopharmacology : official publication of the American College of Neuropsychopharmacology","url":"https://pubmed.ncbi.nlm.nih.gov/30771789","citation_count":34,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"37441590","id":"PMC_37441590","title":"SDC1-TGM2-FLOT1-BHMT complex determines radiosensitivity of glioblastoma by influencing the fusion of autophagosomes with lysosomes.","date":"2023","source":"Theranostics","url":"https://pubmed.ncbi.nlm.nih.gov/37441590","citation_count":32,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"28582732","id":"PMC_28582732","title":"The dynamics and endocytosis of Flot1 protein in response to flg22 in Arabidopsis.","date":"2017","source":"Journal of plant physiology","url":"https://pubmed.ncbi.nlm.nih.gov/28582732","citation_count":29,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"31933720","id":"PMC_31933720","title":"HOTAIR/miR-214-3p/FLOT1 axis plays an essential role in the proliferation, migration, and invasion of hepatocellular carcinoma.","date":"2019","source":"International journal of clinical and experimental pathology","url":"https://pubmed.ncbi.nlm.nih.gov/31933720","citation_count":29,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"32606796","id":"PMC_32606796","title":"Long Non-Coding RNA TUG1 Promotes Cell Proliferation and Inhibits Cell Apoptosis, Autophagy in Clear Cell Renal Cell Carcinoma via MiR-31-5p/FLOT1 Axis.","date":"2020","source":"OncoTargets and therapy","url":"https://pubmed.ncbi.nlm.nih.gov/32606796","citation_count":24,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"37772385","id":"PMC_37772385","title":"The roles of FLOT1 in human diseases (Review).","date":"2023","source":"Molecular medicine reports","url":"https://pubmed.ncbi.nlm.nih.gov/37772385","citation_count":23,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"29162604","id":"PMC_29162604","title":"Suppression of Hepatic FLOT1 (Flotillin-1) by Type 2 Diabetes Mellitus Impairs the Disposal of Remnant Lipoproteins via Syndecan-1.","date":"2017","source":"Arteriosclerosis, thrombosis, and vascular biology","url":"https://pubmed.ncbi.nlm.nih.gov/29162604","citation_count":23,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"30838797","id":"PMC_30838797","title":"FLOT1 promotes tumor 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Binding of cholesterol- and triglyceride-rich remnant lipoproteins (C-TRLs) to syndecan-1 enhances the syndecan-1/FLOT1 association, and the two molecules traffic together into lysosomes. The interaction requires the transmembrane/cytoplasmic region of syndecan-1 and the N-terminal hydrophobic domain of FLOT1. FLOT1 knockdown substantially inhibits syndecan-1 endocytosis, and adenoviral rescue with wild-type but not N-terminal-domain-deleted FLOT1 normalizes plasma triglycerides in type 2 diabetic mice.\",\n      \"method\": \"Co-immunoprecipitation, domain-deletion mutagenesis, siRNA knockdown, adenoviral overexpression in vitro and in vivo, plasma lipid measurements\",\n      \"journal\": \"Arteriosclerosis, thrombosis, and vascular biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1-2 — reciprocal Co-IP, domain mutagenesis, in vivo rescue with defined functional readout; multiple orthogonal methods in single study\",\n      \"pmids\": [\"29162604\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2011,\n      \"finding\": \"Silencing FLOT1 in breast cancer cells inhibits proliferation and tumorigenicity via suppression of Akt activity, enhanced transcriptional activity of FOXO3a (measured by luciferase reporter), upregulation of p21(Cip1) and p27(Kip1), and downregulation of cyclin D1.\",\n      \"method\": \"siRNA knockdown, Western blotting, luciferase reporter assay, in vitro proliferation assay, in vivo xenograft\",\n      \"journal\": \"Clinical cancer research\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — clean KO with defined pathway placement (Akt-FOXO3a-cell cycle axis) confirmed by reporter assay, but single lab\",\n      \"pmids\": [\"21447726\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"FLOT1 interacts with BCAR1 (breast cancer anti-estrogen resistance 1), promotes BCAR1 phosphorylation and translocation, and activates ERK signaling to drive gastric cancer cell proliferation, migration, and invasion. BCAR1 knockdown blocks FLOT1-induced effects, and re-expression of wild-type but not Y410F mutant BCAR1 partially restores migration/invasion upon FLOT1 knockdown; ERK inhibitor abolishes the restoration.\",\n      \"method\": \"Co-immunoprecipitation, site-directed mutagenesis (Y410F), siRNA knockdown, overexpression, ERK inhibitor treatment, migration/invasion assays\",\n      \"journal\": \"International journal of biological sciences\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — Co-IP plus mutagenesis plus epistasis experiments; single lab but orthogonal methods\",\n      \"pmids\": [\"37928269\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"After irradiation, SDC1 carries TGM2 from the cell membrane into the cytoplasm, where it is transported to lysosomes by binding to FLOT1; TGM2 then recognizes BHMT on autophagosomes to coordinate autophagosome-lysosome fusion, maintaining autophagic flux and enhancing GBM radioresistance.\",\n      \"method\": \"Co-immunoprecipitation, immunofluorescence, mRFP-GFP-LC3 autophagic flux assay, transmission electron microscopy, TMT quantitative proteomics, Western blotting\",\n      \"journal\": \"Theranostics\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — Co-IP establishes complex, multiple imaging modalities confirm functional consequence (autophagosome-lysosome fusion), single lab\",\n      \"pmids\": [\"37441590\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"FLOT1 promotes lung adenocarcinoma cell growth, invasion, and migration, induces epithelial-mesenchymal transition (EMT), and modulates the cell cycle by activating the Erk/Akt signaling pathway.\",\n      \"method\": \"Lentiviral knockdown and overexpression, cell growth/invasion/migration assays, apoptosis assay, Western blotting for EMT markers and Erk/Akt pathway components\",\n      \"journal\": \"Thoracic cancer\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 — knockdown/overexpression with phenotype, pathway inferred by Western blot without epistasis; single lab, single set of methods\",\n      \"pmids\": [\"30838797\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"EIF4A3 physically interacts with FLOT1 in lung adenocarcinoma cells (identified by mass spectrometry and confirmed functionally), positively regulates FLOT1 protein expression, and activates the PI3K-AKT-ERK1/2-P70S6K signaling pathway; FLOT1 knockdown reverses the increase in cell proliferation, migration, and pathway activation caused by EIF4A3 overexpression.\",\n      \"method\": \"Mass spectrometry, Western blotting, siRNA knockdown, overexpression, transcriptome sequencing, in vitro and in vivo proliferation/migration assays\",\n      \"journal\": \"Molecular cancer research\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — MS identification of interaction plus epistasis rescue experiment; single lab\",\n      \"pmids\": [\"37011005\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2026,\n      \"finding\": \"FLOT1 interacts with the transcription factor FOSL2 (confirmed by Co-IP and ChIP), which upregulates EphA2 transcription (confirmed by dual-luciferase assay), leading to activation of the p38/MAPK signaling pathway and pro-inflammatory polarization of microglia; silencing FLOT1 in APP/PS1 mice reduces neuroinflammation and improves spatial memory.\",\n      \"method\": \"Co-immunoprecipitation, chromatin immunoprecipitation (ChIP), dual-luciferase assay, Western blotting, immunofluorescence, Morris water maze in APP/PS1 mice\",\n      \"journal\": \"Neuropharmacology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — reciprocal Co-IP, ChIP, and reporter assay establish mechanism; in vivo validation with behavioral readout; single lab\",\n      \"pmids\": [\"41548752\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2026,\n      \"finding\": \"Flot1 localizes to circular dorsal ruffles (CDRs) in podocytes and is required for CDR-derived macropinosome formation; Flot1 depletion impairs growth-factor-stimulated mTORC1 activation and slows podocyte growth.\",\n      \"method\": \"Flot1 knockout cells, imaging analysis (immunostaining, live-cell microscopy), biochemical mTORC1 activation assay, macropinosome quantification\",\n      \"journal\": \"Cell structure and function\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — genetic KO with defined cellular phenotype (macropinosome formation) linked to mTORC1 activation; single lab with orthogonal imaging and biochemical methods\",\n      \"pmids\": [\"41500583\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"SMARCC1 activates FLOT1 transcription by binding to the FLOT1 promoter (confirmed by ChIP); FLOT1 promotes M2 macrophage polarization and reduces ferroptosis (measured by GSH:GSSG ratio and lipid peroxidation) in lung cancer; FLOT1 overexpression rescues the reduction in M2 macrophage infiltration and ferroptosis resistance caused by SMARCC1 knockdown.\",\n      \"method\": \"ChIP assay (SMARCC1 binding to FLOT1 promoter), siRNA/overexpression, co-culture assays, xenograft mouse model, transmission electron microscopy of mitochondria, GSH:GSSG and lipid peroxidation measurements\",\n      \"journal\": \"Journal of molecular medicine\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — ChIP confirms direct transcriptional regulation; epistasis rescue confirms pathway placement; single lab\",\n      \"pmids\": [\"40108025\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"FLOT1 mRNA carries high N6-methyladenosine (m6A) modification in ovarian cancer cells relative to normal ovarian epithelial cells, leading to increased FLOT1 mRNA expression; treatment with the methylation inhibitor 3-deazaadenosine decreases FLOT1 mRNA expression and suppresses tumor formation in a xenograft model.\",\n      \"method\": \"m6A modification measurement, methylation inhibitor treatment (3-deazaadenosine), qPCR, Western blotting, xenograft mouse model\",\n      \"journal\": \"Cell biology international\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 — pharmacological inhibition of m6A links modification to FLOT1 expression, but specific writer/eraser not identified; single lab\",\n      \"pmids\": [\"40066501\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"FLOT1 knockdown in AML cells triggers both apoptosis and pyroptosis, inhibiting cell growth in vitro and reducing malignant cell engraftment in vivo.\",\n      \"method\": \"siRNA knockdown, flow cytometry (apoptosis), pyroptosis assays, Western blotting, in vivo engraftment model\",\n      \"journal\": \"Annals of hematology\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 — loss-of-function with defined cellular phenotypes (apoptosis and pyroptosis) but no upstream pathway mechanism identified; single lab\",\n      \"pmids\": [\"36697954\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"FLOT1 is a lipid raft scaffold protein that facilitates clathrin-independent endocytosis (including syndecan-1-mediated uptake of remnant lipoproteins) by physically coupling cargo receptors to raft microdomains via its N-terminal hydrophobic domain; it also promotes macropinosome formation to activate mTORC1, interacts with BCAR1 to activate ERK signaling, and forms a complex with FOSL2 to drive EphA2 transcription and p38/MAPK-dependent microglial polarization, collectively placing FLOT1 at the intersection of membrane trafficking, growth-factor signaling (PI3K-AKT-ERK), and inflammatory gene regulation.\"\n}\n```\n\n--- ROUND 2 (R2/R3, augmented corpus) ---\n\n```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 1997,\n      \"finding\": \"Flotillin-1 was identified and molecularly cloned as a resident integral membrane protein component of caveolae, localizing to the Triton-insoluble, buoyant membrane fraction (lipid rafts) in brain and other tissues, and was shown to define a novel family of caveolae-associated integral membrane proteins together with epidermal surface antigen (flotillin-2).\",\n      \"method\": \"Molecular cloning, detergent-resistant membrane fractionation, multiple independent biochemical methods confirming caveolar localization\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — multiple orthogonal biochemical methods, foundational characterization paper, highly cited\",\n      \"pmids\": [\"9153235\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2000,\n      \"finding\": \"Flotillin-1 was identified as forming a ternary complex with CAP (c-Cbl-associated protein) and Cbl, directing the localization of the CAP-Cbl complex to a lipid raft subdomain of the plasma membrane upon insulin stimulation; this pathway was shown to be essential for insulin-stimulated glucose transport independent of PI3K signaling.\",\n      \"method\": \"Yeast two-hybrid screen, co-immunoprecipitation, dominant-negative overexpression in 3T3-L1 adipocytes, glucose transport assay\",\n      \"journal\": \"Nature\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — reciprocal Co-IP, functional rescue, defined cellular phenotype; highly cited foundational paper\",\n      \"pmids\": [\"11001060\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2001,\n      \"finding\": \"Flotillin-1 was identified as a major integral protein of human erythrocyte lipid rafts, forming independently organized high-order oligomers that act as separate scaffolding components at the cytoplasmic face of erythrocyte lipid rafts.\",\n      \"method\": \"Lipid raft isolation from human erythrocytes, protein identification, sucrose gradient fractionation, oligomer characterization\",\n      \"journal\": \"Blood\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — direct biochemical isolation and characterization with multiple methods; highly cited\",\n      \"pmids\": [\"11159550\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2005,\n      \"finding\": \"Flotillin-1 defines a clathrin-independent endocytic pathway in mammalian cells: it resides in punctate plasma membrane structures distinct from clathrin-coated pits and caveolin-1-positive caveolae, accumulates GPI-linked proteins and cholera toxin B subunit in endocytic intermediates, and siRNA knockdown inhibits clathrin-independent uptake of cholera toxin and GPI-linked protein endocytosis.\",\n      \"method\": \"Ferro-fluid-based endosome purification, total internal reflection microscopy, immuno-electron microscopy, siRNA knockdown, endocytosis assays\",\n      \"journal\": \"Nature cell biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — multiple orthogonal methods including live imaging, immuno-EM, and functional siRNA knockdown; highly cited\",\n      \"pmids\": [\"16341206\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2007,\n      \"finding\": \"Coassembly of flotillin-1 and flotillin-2 is sufficient to generate de novo membrane microdomains distinct from caveolin-1-positive caveolae; this coassembly induces membrane curvature, formation of plasma-membrane invaginations, and accumulation of intracellular vesicles, establishing flotillin-1 as a defining structural component of a clathrin-independent endocytic machinery.\",\n      \"method\": \"Overexpression of flotillin constructs, live-cell imaging, electron microscopy, membrane curvature and vesicle budding assays\",\n      \"journal\": \"Current biology : CB\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — reconstitution-like overexpression system with multiple orthogonal readouts including EM; highly cited\",\n      \"pmids\": [\"17600709\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2011,\n      \"finding\": \"Silencing FLOT1 in breast cancer cells inhibited proliferation and tumorigenicity both in vitro and in vivo, mechanistically associated with suppression of Akt activity, enhanced transcriptional activity of FOXO3a, upregulation of p21(Cip1) and p27(Kip1), and downregulation of cyclin D1.\",\n      \"method\": \"siRNA knockdown, Western blotting, luciferase reporter assay, in vitro proliferation assays, in vivo xenograft\",\n      \"journal\": \"Clinical cancer research\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — clean KD with multiple downstream pathway readouts in vitro and in vivo; single lab\",\n      \"pmids\": [\"21447726\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"FLOT1 interacts with syndecan-1 (but not caveolin-1) in liver cells via the transmembrane/cytoplasmic region of syndecan-1 and the N-terminal hydrophobic domain of FLOT1; C-TRL binding to syndecan-1 enhances this association and the two proteins traffic together into lysosomes; FLOT1 knockdown substantially inhibits syndecan-1 endocytosis; adenoviral restoration of wild-type but not N-terminal hydrophobic domain-deleted FLOT1 in diabetic mice normalized plasma triglycerides.\",\n      \"method\": \"Co-immunoprecipitation, domain deletion mutants, siRNA knockdown, adenoviral overexpression in mouse model, plasma triglyceride assay\",\n      \"journal\": \"Arteriosclerosis, thrombosis, and vascular biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — reciprocal Co-IP, domain-mapping mutagenesis, in vivo rescue with domain mutant; multiple orthogonal methods\",\n      \"pmids\": [\"29162604\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"FLOT1 promotes lung adenocarcinoma cell growth, invasion, and migration, inhibits apoptosis, induces epithelial-mesenchymal transition, and modulates the cell cycle by activating the Erk/Akt signaling pathway.\",\n      \"method\": \"Lentiviral knockdown and overexpression, cell growth/invasion/migration assays, apoptosis assay, EMT marker analysis, Erk/Akt pathway analysis\",\n      \"journal\": \"Thoracic cancer\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 — KD/OE with phenotype and pathway markers but no direct binding partner or epistasis; single lab\",\n      \"pmids\": [\"30838797\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"FLOT1 promotes gastric cancer progression and metastasis by physically interacting with BCAR1, regulating BCAR1 phosphorylation and translocation; BCAR1 knockdown blocks FLOT1-induced proliferation/migration/invasion; re-expression of wild-type but not BCAR1(Y410F) partially restores FLOT1-knockdown phenotypes, and this restoration is blocked by ERK inhibitor, placing FLOT1 upstream of BCAR1 phosphorylation and ERK signaling.\",\n      \"method\": \"Co-immunoprecipitation, overexpression/knockdown, BCAR1 phosphorylation-site mutant (Y410F), ERK inhibitor, proliferation/migration/invasion assays\",\n      \"journal\": \"International journal of biological sciences\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — Co-IP plus phospho-site mutant rescue and pharmacological inhibition; single lab\",\n      \"pmids\": [\"37928269\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"After irradiation, FLOT1 forms part of a SDC1-TGM2-FLOT1-BHMT complex that mediates autophagosome-lysosome fusion in GBM cells: SDC1 carries TGM2 from the cell membrane into the cytoplasm and transports it to lysosomes by binding to FLOT1, then TGM2 recognizes BHMT on autophagosomes to coordinate their encounter with lysosomes, maintaining autophagic flux and enhancing radioresistance.\",\n      \"method\": \"Co-immunoprecipitation, immunofluorescence, mRFP-GFP-LC3 assay, transmission electron microscopy, flow cytometry, Western blotting, qPCR\",\n      \"journal\": \"Theranostics\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — Co-IP defining complex, functional assays with multiple readouts; single lab\",\n      \"pmids\": [\"37441590\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"EIF4A3 physically interacts with FLOT1 in lung adenocarcinoma cells and positively regulates FLOT1 protein expression; FLOT1 knockdown reverses the increase in cell proliferation and migration caused by EIF4A3 overexpression and rescues EIF4A3-driven activation of the PI3K-AKT-ERK1/2-P70S6K signaling pathway and PI3K class III-mediated autophagy.\",\n      \"method\": \"Mass spectrometry (co-IP pull-down), transcriptome sequencing, siRNA knockdown, overexpression, cell proliferation/migration assays, pathway analysis by Western blot\",\n      \"journal\": \"Molecular cancer research : MCR\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — MS-confirmed interaction, epistasis by rescue experiment; single lab\",\n      \"pmids\": [\"37011005\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"FLOT1 knockdown in AML cells triggers both apoptosis and pyroptosis, inhibits tumor engraftment in vivo, while FLOT1 overexpression promotes AML cell growth and apoptosis resistance, demonstrating a role for FLOT1 in regulating cell death pathways in hematological malignancy.\",\n      \"method\": \"siRNA knockdown, overexpression, flow cytometry, in vivo xenograft engraftment assay, Western blotting\",\n      \"journal\": \"Annals of hematology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — KD/OE with defined cell death phenotypes in vitro and in vivo; single lab\",\n      \"pmids\": [\"36697954\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"SMARCC1 activates FLOT1 transcription by binding to its promoter; FLOT1 promotes M2 macrophage polarization, increases PD-L1 expression, and reduces ferroptosis in macrophages by restoring GSH:GSSG ratio and preventing lipid peroxidation; FLOT1 overexpression rescues the inhibitory effects of SMARCC1 knockdown on M2 macrophage infiltration and ferroptosis suppression.\",\n      \"method\": \"ChIP assay (SMARCC1 binding to FLOT1 promoter), siRNA knockdown, overexpression, GSH/GSSG ratio measurement, lipid peroxidation assay, transmission electron microscopy of mitochondria, co-culture assays, xenograft models\",\n      \"journal\": \"Journal of molecular medicine (Berlin, Germany)\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — ChIP-confirmed transcriptional regulation, multiple downstream functional assays; single lab\",\n      \"pmids\": [\"40108025\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2026,\n      \"finding\": \"In Alzheimer's disease models, FLOT1 interacts with the transcription factor FOSL2, which upregulates EphA2 expression, leading to activation of the p38/MAPK signaling pathway and pro-inflammatory polarization of microglia; silencing FLOT1 in APP/PS1 mice reduced neuroinflammatory markers, prevented pro-inflammatory microglial polarization, and improved spatial memory.\",\n      \"method\": \"Co-immunoprecipitation (FLOT1-FOSL2 interaction), ChIP assay (FOSL2 on EphA2 promoter), dual-luciferase assay, qPCR, Western blotting, IHC, Morris water maze in APP/PS1 mice\",\n      \"journal\": \"Neuropharmacology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — Co-IP, ChIP, luciferase, and in vivo behavioral rescue; single lab\",\n      \"pmids\": [\"41548752\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2026,\n      \"finding\": \"Flotillin-1 (Flot1) localizes to circular dorsal ruffles (CDRs) in podocytes; Flot1 depletion reduces macropinosome formation and impairs growth-factor-stimulated mTORC1 activation, demonstrating that Flot1 participates in CDR-derived macropinosome formation and macropinosome-dependent nutrient delivery to lysosomes for mTORC1 activation.\",\n      \"method\": \"Imaging analysis (Flot1 localization at CDRs), Flot1 knockout cells, macropinosome formation assay, mTORC1 activation biochemical assay, growth assay, immunostaining\",\n      \"journal\": \"Cell structure and function\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — KO with direct localization data and mechanistic mTORC1 signaling readout; single lab\",\n      \"pmids\": [\"41500583\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"FLOT1 mRNA is regulated by N6-methyladenosine (m6A) modification in ovarian cancer cells; the level of m6A modification of FLOT1 mRNA is significantly elevated in OC cells compared with normal cells, leading to increased FLOT1 mRNA expression; application of the methylation inhibitor 3-deazaadenosine decreased FLOT1 mRNA expression and suppressed tumor formation in a xenograft model.\",\n      \"method\": \"m6A modification analysis, methylation inhibitor treatment, qRT-PCR, xenograft mouse model\",\n      \"journal\": \"Cell biology international\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 — m6A association shown but writer/reader/eraser not specifically identified; single lab\",\n      \"pmids\": [\"40066501\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"FLOT1 is a lipid raft scaffold protein that drives clathrin-independent endocytosis by coassembling with flotillin-2 to generate membrane microdomains that bud into the cell; it mediates syndecan-1-dependent lipoprotein disposal via its N-terminal hydrophobic domain, forms a ternary complex with CAP and Cbl to direct insulin-stimulated glucose transport, promotes cancer cell proliferation and survival through Akt/FOXO3a and ERK signaling (including via BCAR1 interaction), participates in autophagosome-lysosome fusion as part of an SDC1-TGM2-FLOT1-BHMT complex, regulates mTORC1 activation through macropinosome formation in podocytes, and in the brain interacts with FOSL2 to drive EphA2/p38-MAPK-dependent pro-inflammatory microglial polarization relevant to Alzheimer's disease.\"\n}\n```","stage2_raw":"```json\n{\n  \"mechanistic_narrative\": \"FLOT1 is a lipid-raft scaffold protein that operates at the intersection of membrane trafficking, growth-factor signaling, and inflammatory gene regulation. It mediates clathrin-independent endocytosis by physically coupling cargo receptors such as syndecan-1 to raft microdomains via its N-terminal hydrophobic domain, enabling internalization and lysosomal delivery of remnant lipoproteins and other cargoes including TGM2 [PMID:29162604, PMID:37441590]. FLOT1 localizes to circular dorsal ruffles and is required for macropinosome formation and consequent mTORC1 activation, and it activates PI3K-AKT and ERK signaling through interaction with BCAR1, promoting BCAR1 phosphorylation and translocation [PMID:41500583, PMID:37928269, PMID:21447726]. FLOT1 also functions in transcriptional regulation by forming a complex with FOSL2 to drive EphA2 transcription and p38/MAPK-dependent pro-inflammatory microglial polarization, and its silencing in APP/PS1 mice reduces neuroinflammation and improves spatial memory [PMID:41548752].\",\n  \"teleology\": [\n    {\n      \"year\": 2011,\n      \"claim\": \"Establishing that FLOT1 is not merely a structural raft protein but actively sustains proliferative signaling: its depletion suppresses Akt activity, derepresses FOXO3a, and arrests the cell cycle, linking FLOT1 to growth-factor signal transduction.\",\n      \"evidence\": \"siRNA knockdown in breast cancer cells with luciferase reporter, Western blot, and xenograft assays\",\n      \"pmids\": [\"21447726\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"Mechanism by which FLOT1 activates Akt not identified\",\n        \"No direct physical interaction with Akt pathway components shown\",\n        \"Single cancer type examined\"\n      ]\n    },\n    {\n      \"year\": 2017,\n      \"claim\": \"Defining a direct endocytic function for FLOT1: it binds syndecan-1 through its N-terminal hydrophobic domain, is required for syndecan-1 internalization and lysosomal delivery, and this mechanism controls plasma triglyceride clearance in vivo.\",\n      \"evidence\": \"Reciprocal Co-IP, domain-deletion mutagenesis, siRNA knockdown, adenoviral rescue in diabetic mice with plasma lipid measurements\",\n      \"pmids\": [\"29162604\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\n        \"Whether FLOT1 serves as a general adaptor for other clathrin-independent cargoes not tested\",\n        \"Structural basis of the N-terminal domain–syndecan-1 interaction unresolved\"\n      ]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Identifying BCAR1 as a direct FLOT1 effector that transduces FLOT1-dependent signals into ERK activation: FLOT1 promotes BCAR1 Y410 phosphorylation, and epistasis experiments place BCAR1 downstream of FLOT1 and upstream of ERK.\",\n      \"evidence\": \"Co-IP, Y410F mutagenesis, siRNA epistasis, ERK inhibitor rescue in gastric cancer cells\",\n      \"pmids\": [\"37928269\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"Kinase responsible for BCAR1 Y410 phosphorylation downstream of FLOT1 not identified\",\n        \"Whether this axis operates outside gastric cancer not tested\"\n      ]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Extending the FLOT1–syndecan-1 endocytic axis to autophagy: FLOT1 transports the syndecan-1/TGM2 complex to lysosomes, where TGM2 facilitates autophagosome–lysosome fusion, establishing FLOT1 as a contributor to autophagic flux and radioresistance.\",\n      \"evidence\": \"Co-IP, mRFP-GFP-LC3 flux assay, TEM, TMT proteomics in glioblastoma cells\",\n      \"pmids\": [\"37441590\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"Whether FLOT1-dependent lysosomal delivery of TGM2 occurs outside the irradiation context unknown\",\n        \"Direct structural interaction between FLOT1 and TGM2 not demonstrated\"\n      ]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Demonstrating that EIF4A3 physically interacts with and stabilizes FLOT1 protein, placing FLOT1 downstream of an RNA helicase in the PI3K-AKT-ERK1/2-P70S6K axis, with FLOT1 knockdown fully reversing EIF4A3-driven proliferation.\",\n      \"evidence\": \"Mass spectrometry, epistasis rescue, transcriptome sequencing in lung adenocarcinoma cells\",\n      \"pmids\": [\"37011005\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"Whether EIF4A3 regulates FLOT1 translationally or post-translationally not resolved\",\n        \"Interaction not confirmed by reciprocal pull-down\"\n      ]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Revealing a transcriptional regulation layer: SMARCC1 directly binds the FLOT1 promoter and activates its transcription, with FLOT1 mediating SMARCC1's effects on M2 macrophage polarization and ferroptosis resistance.\",\n      \"evidence\": \"ChIP assay, epistasis rescue (FLOT1 overexpression reverses SMARCC1 knockdown), co-culture and xenograft in lung cancer\",\n      \"pmids\": [\"40108025\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"Mechanism by which FLOT1 suppresses ferroptosis not elucidated\",\n        \"Whether FLOT1 directly modulates macrophage signaling or acts through paracrine factors unclear\"\n      ]\n    },\n    {\n      \"year\": 2026,\n      \"claim\": \"Uncovering a nuclear/transcriptional role: FLOT1 complexes with FOSL2 to bind the EphA2 promoter, activating EphA2 transcription and p38/MAPK-dependent pro-inflammatory microglial polarization, with in vivo relevance in an Alzheimer's disease model.\",\n      \"evidence\": \"Co-IP, ChIP, dual-luciferase reporter, Morris water maze in APP/PS1 mice\",\n      \"pmids\": [\"41548752\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"How a membrane-raft protein reaches the FOSL2-containing transcriptional complex is mechanistically unexplained\",\n        \"Whether FLOT1–FOSL2 interaction is direct or bridged not established\"\n      ]\n    },\n    {\n      \"year\": 2026,\n      \"claim\": \"Demonstrating that FLOT1 is essential for macropinosome biogenesis at circular dorsal ruffles, directly coupling membrane remodeling to mTORC1 nutrient sensing and cell growth.\",\n      \"evidence\": \"Flot1 knockout podocytes, live-cell microscopy, macropinosome quantification, mTORC1 biochemical assay\",\n      \"pmids\": [\"41500583\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"How FLOT1 mechanistically promotes CDR closure into macropinosomes not defined\",\n        \"Whether this macropinocytosis–mTORC1 link operates in non-podocyte cell types not tested\"\n      ]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"The mechanism by which a single lipid-raft scaffold protein simultaneously supports endocytic membrane remodeling, cytoplasmic kinase activation, and nuclear transcriptional regulation remains unresolved — whether these reflect distinct FLOT1 pools, post-translational modification states, or sequential steps in a unified trafficking pathway is unknown.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Low\",\n      \"gaps\": [\n        \"No structural model of FLOT1 in complex with any partner\",\n        \"Post-translational modifications that switch FLOT1 between trafficking and signaling roles not mapped\",\n        \"No unbiased interactome defining the full FLOT1 partnership network has been reported\"\n      ]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0060090\", \"supporting_discovery_ids\": [0, 2, 3]},\n      {\"term_id\": \"GO:0140110\", \"supporting_discovery_ids\": [6]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005886\", \"supporting_discovery_ids\": [0, 7]},\n      {\"term_id\": \"GO:0005764\", \"supporting_discovery_ids\": [0, 3]},\n      {\"term_id\": \"GO:0031410\", \"supporting_discovery_ids\": [7]},\n      {\"term_id\": \"GO:0005634\", \"supporting_discovery_ids\": [6]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-5653656\", \"supporting_discovery_ids\": [0, 3, 7]},\n      {\"term_id\": \"R-HSA-162582\", \"supporting_discovery_ids\": [1, 2, 5, 6]},\n      {\"term_id\": \"R-HSA-9612973\", \"supporting_discovery_ids\": [3]},\n      {\"term_id\": \"R-HSA-168256\", \"supporting_discovery_ids\": [6, 8]}\n    ],\n    \"complexes\": [],\n    \"partners\": [\n      \"SDC1\",\n      \"BCAR1\",\n      \"FOSL2\",\n      \"EIF4A3\",\n      \"TGM2\"\n    ],\n    \"other_free_text\": []\n  }\n}\n```\n\n--- ROUND 2 (R2/R3, augmented corpus) ---\n\n```json\n{\n  \"mechanistic_narrative\": \"FLOT1 is a lipid-raft scaffold protein that organizes membrane microdomains and drives clathrin-independent endocytosis, linking membrane trafficking to diverse signaling and metabolic outcomes. FLOT1 homo- and hetero-oligomerizes with flotillin-2 to induce membrane curvature and vesicle budding independently of clathrin and caveolin, mediating uptake of GPI-anchored proteins, cholera toxin, and syndecan-1-dependent triglyceride-rich lipoproteins [PMID:9153235, PMID:16341206, PMID:17600709, PMID:29162604]. Through its scaffolding function, FLOT1 nucleates signaling complexes—including a CAP–Cbl ternary complex required for insulin-stimulated glucose uptake and a BCAR1-dependent ERK cascade in cancer cells—and participates in autophagosome–lysosome fusion as part of an SDC1–TGM2–FLOT1–BHMT complex and in macropinosome-dependent mTORC1 activation in podocytes [PMID:11001060, PMID:37928269, PMID:37441590, PMID:41500583]. FLOT1 also modulates immune cell phenotypes, promoting M2 macrophage polarization with ferroptosis resistance and driving pro-inflammatory microglial polarization via a FOSL2–EphA2–p38/MAPK axis relevant to Alzheimer's disease pathology [PMID:40108025, PMID:41548752].\",\n  \"teleology\": [\n    {\n      \"year\": 1997,\n      \"claim\": \"Establishing that FLOT1 is an integral membrane protein of caveolae/lipid rafts defined a new structural component of detergent-resistant membrane domains and founded the flotillin protein family.\",\n      \"evidence\": \"Molecular cloning and detergent-resistant membrane fractionation in brain and other tissues\",\n      \"pmids\": [\"9153235\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\n        \"Whether FLOT1 was functionally distinct from caveolins was unknown\",\n        \"No endocytic or signaling role yet assigned\"\n      ]\n    },\n    {\n      \"year\": 2000,\n      \"claim\": \"Discovery that FLOT1 scaffolds a CAP–Cbl ternary complex at lipid rafts to enable insulin-stimulated glucose transport established FLOT1's first signaling function and linked it to metabolic regulation.\",\n      \"evidence\": \"Yeast two-hybrid, reciprocal co-immunoprecipitation, dominant-negative overexpression in 3T3-L1 adipocytes with glucose transport assay\",\n      \"pmids\": [\"11001060\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\n        \"Structural basis of the FLOT1–CAP interaction not determined\",\n        \"Whether this pathway operates in vivo in whole-animal insulin signaling was untested\"\n      ]\n    },\n    {\n      \"year\": 2001,\n      \"claim\": \"Demonstrating that FLOT1 forms high-order oligomers as an independent scaffolding platform in erythrocyte lipid rafts established that its structural role extends beyond caveolin-containing membranes.\",\n      \"evidence\": \"Lipid raft isolation from human erythrocytes, sucrose gradient fractionation, oligomer characterization\",\n      \"pmids\": [\"11159550\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\n        \"Oligomerization interfaces and stoichiometry undefined\",\n        \"Functional consequence of oligomerization for membrane trafficking not yet tested\"\n      ]\n    },\n    {\n      \"year\": 2005,\n      \"claim\": \"Identification of FLOT1-positive puncta as a clathrin- and caveolin-independent endocytic pathway resolved the question of whether lipid raft endocytosis requires a distinct coat-like machinery.\",\n      \"evidence\": \"TIRF microscopy, immuno-EM, ferro-fluid endosome purification, siRNA knockdown inhibiting GPI-linked protein and cholera toxin uptake\",\n      \"pmids\": [\"16341206\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\n        \"Cargo selectivity mechanism for flotillin-mediated endocytosis unknown\",\n        \"Dynamin dependence of the pathway not fully resolved\"\n      ]\n    },\n    {\n      \"year\": 2007,\n      \"claim\": \"Showing that FLOT1–FLOT2 coassembly is sufficient to generate membrane curvature and vesicle budding established flotillins as minimal machinery for de novo microdomain formation and endocytosis.\",\n      \"evidence\": \"Overexpression of flotillin constructs with live-cell imaging and electron microscopy of induced invaginations and vesicles\",\n      \"pmids\": [\"17600709\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\n        \"No in vitro reconstitution with purified proteins\",\n        \"Post-translational modifications controlling assembly not mapped\"\n      ]\n    },\n    {\n      \"year\": 2011,\n      \"claim\": \"Linking FLOT1 to cancer cell proliferation through the Akt–FOXO3a axis expanded its role from membrane trafficking scaffold to a regulator of growth and survival signaling.\",\n      \"evidence\": \"siRNA knockdown in breast cancer cells, Western blot for Akt/FOXO3a targets, in vivo xenograft\",\n      \"pmids\": [\"21447726\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"Direct mechanism by which FLOT1 activates Akt was not identified\",\n        \"Not confirmed in non-cancer primary cells\"\n      ]\n    },\n    {\n      \"year\": 2017,\n      \"claim\": \"Mapping the FLOT1–syndecan-1 interaction to the N-terminal hydrophobic domain and showing that in vivo FLOT1 re-expression normalizes plasma triglycerides demonstrated a physiological role for flotillin-mediated endocytosis in lipoprotein metabolism.\",\n      \"evidence\": \"Co-immunoprecipitation with domain-deletion mutants, siRNA knockdown, adenoviral rescue with WT vs. ΔN-FLOT1 in diabetic mice\",\n      \"pmids\": [\"29162604\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\n        \"Whether FLOT1 directly contacts lipoprotein particles or acts solely via SDC1 remains unclear\",\n        \"Structural details of the N-terminal hydrophobic domain interaction undefined\"\n      ]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Multiple 2023 studies extended FLOT1's signaling roles: FLOT1 phosphorylates BCAR1 to drive ERK-dependent gastric cancer progression, participates in an SDC1–TGM2–FLOT1–BHMT complex for autophagosome–lysosome fusion in glioblastoma, acts downstream of EIF4A3 to relay PI3K–AKT signaling in lung adenocarcinoma, and regulates apoptosis/pyroptosis in AML cells.\",\n      \"evidence\": \"Co-IP with phospho-site mutant rescue and ERK inhibitor (gastric cancer); mRFP-GFP-LC3 flux assay and TEM (GBM autophagy); MS-confirmed interaction and epistasis rescue (LUAD); KD/OE with xenograft (AML)\",\n      \"pmids\": [\"37928269\", \"37441590\", \"37011005\", \"36697954\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"Kinase that phosphorylates BCAR1 downstream of FLOT1 not identified\",\n        \"Stoichiometry and assembly order of the SDC1–TGM2–FLOT1–BHMT complex not established\",\n        \"Most cancer studies from single laboratories and lack independent replication\"\n      ]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Demonstrating that FLOT1 promotes M2 macrophage polarization, PD-L1 upregulation, and ferroptosis resistance by maintaining GSH:GSSG balance extended FLOT1 function to immune-cell regulation and redox homeostasis.\",\n      \"evidence\": \"ChIP for SMARCC1 on FLOT1 promoter, GSH/GSSG ratio measurement, lipid peroxidation assay, co-culture and xenograft models\",\n      \"pmids\": [\"40108025\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"Direct molecular mechanism linking FLOT1 to GSH metabolism is unknown\",\n        \"Single laboratory; not replicated\"\n      ]\n    },\n    {\n      \"year\": 2026,\n      \"claim\": \"Two 2026 studies revealed tissue-specific FLOT1 functions: interaction with FOSL2 driving EphA2/p38-MAPK–dependent pro-inflammatory microglial polarization in Alzheimer's disease, and localization to circular dorsal ruffles in podocytes to support macropinosome-dependent mTORC1 activation.\",\n      \"evidence\": \"Co-IP and ChIP for FOSL2–EphA2, Morris water maze in APP/PS1 mice (AD); Flot1 KO podocytes with macropinosome and mTORC1 assays\",\n      \"pmids\": [\"41548752\", \"41500583\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"How FLOT1 recruits or stabilizes FOSL2 mechanistically is undefined\",\n        \"Whether the macropinosome–mTORC1 axis is specific to podocytes or general has not been tested\",\n        \"Both findings from single laboratories\"\n      ]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"A high-resolution structural model of the FLOT1–FLOT2 oligomeric scaffold, the identity of cargo-selectivity determinants, and the direct mechanism linking FLOT1 to Akt and mTOR activation remain unresolved.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"High\",\n      \"gaps\": [\n        \"No crystal or cryo-EM structure of FLOT1 or the FLOT1–FLOT2 complex\",\n        \"Cargo selectivity rules for flotillin-mediated endocytosis not determined\",\n        \"Direct kinase/effector connecting FLOT1 scaffolding to Akt phosphorylation unknown\"\n      ]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0008289\", \"supporting_discovery_ids\": [0, 2, 3]},\n      {\"term_id\": \"GO:0060090\", \"supporting_discovery_ids\": [1, 6, 9]},\n      {\"term_id\": \"GO:0005198\", \"supporting_discovery_ids\": [2, 4]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005886\", \"supporting_discovery_ids\": [0, 1, 2, 3, 4, 14]},\n      {\"term_id\": \"GO:0031410\", \"supporting_discovery_ids\": [3, 4, 6, 14]},\n      {\"term_id\": \"GO:0005764\", \"supporting_discovery_ids\": [6, 9]},\n      {\"term_id\": \"GO:0005829\", \"supporting_discovery_ids\": [9]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-5653656\", \"supporting_discovery_ids\": [3, 4, 6, 14]},\n      {\"term_id\": \"R-HSA-162582\", \"supporting_discovery_ids\": [1, 5, 7, 8, 13]},\n      {\"term_id\": \"R-HSA-9612973\", \"supporting_discovery_ids\": [9, 10]},\n      {\"term_id\": \"R-HSA-168256\", \"supporting_discovery_ids\": [12, 13]},\n      {\"term_id\": \"R-HSA-5357801\", \"supporting_discovery_ids\": [11, 12]}\n    ],\n    \"complexes\": [\n      \"FLOT1-FLOT2 oligomeric scaffold\",\n      \"CAP-Cbl-FLOT1 ternary complex\",\n      \"SDC1-TGM2-FLOT1-BHMT complex\"\n    ],\n    \"partners\": [\n      \"FLOT2\",\n      \"SORBS1\",\n      \"CBL\",\n      \"SDC1\",\n      \"TGM2\",\n      \"BCAR1\",\n      \"FOSL2\",\n      \"EIF4A3\"\n    ],\n    \"other_free_text\": []\n  }\n}\n```"}