{"gene":"FLOT2","run_date":"2026-06-09T23:54:43","timeline":{"discoveries":[{"year":2022,"finding":"Flot2 acts as a scaffolding protein that promotes cytoneme-based intercellular transport of Wnt3 in gastric cancer cells. Together with the Wnt co-receptor Ror2, Flot2 determines the number and length of Wnt3-carrying cytonemes. This mechanism is conserved: Flotillins are also necessary for Wnt8a cytonemes during zebrafish embryogenesis.","method":"Live imaging of cytonemes, genetic manipulation of Flot2 and Ror2, zebrafish embryogenesis model, functional proliferation/survival assays","journal":"eLife","confidence":"High","confidence_rationale":"Tier 2 / Moderate — multiple orthogonal methods (imaging, genetic manipulation, in vivo zebrafish model), single lab but mechanistically rigorous with functional validation","pmids":["36040316"],"is_preprint":false},{"year":2021,"finding":"FLOT2 interacts with and stabilizes EphA2 protein in glioma cells; FLOT2 knockdown reduces EphA2 levels, suppresses Akt and NF-κB activities, and induces apoptosis, cell cycle arrest, and EMT inhibition. Restoration of EphA2 rescues the suppressive effects of FLOT2 knockdown, placing FLOT2 upstream of EphA2 in a pro-oncogenic signaling axis.","method":"Co-immunoprecipitation, siRNA knockdown, Western blot, IHC, functional rescue assays","journal":"Biochemical and biophysical research communications","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — reciprocal Co-IP and functional rescue in single lab with multiple orthogonal methods","pmids":["33631676"],"is_preprint":false},{"year":2025,"finding":"LAMP2 preferentially binds FLOT2 in cardiac endothelial cells following IL-4 exposure, and this LAMP2-FLOT2 interaction enhances autophagosome-lysosome fusion. Loss of FLOT2 reverses the LAMP2-mediated rescue of autophagic flux, leading to autophagosome accumulation.","method":"Co-immunoprecipitation, loss-of-function (FLOT2 knockdown), autophagic flux assays (including transmission electron microscopy and LC3/p62 markers), CLP mouse sepsis model","journal":"Autophagy","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — Co-IP, functional KO, multiple orthogonal readouts (TEM, flux markers), single lab","pmids":["40066518"],"is_preprint":false},{"year":2025,"finding":"Flot2 directly interacts with synaptopodin in podocytes and protects it from proteasomal degradation via inhibition of K48-linked polyubiquitination. Podocyte-specific Flot2 knockout worsened albuminuria and podocyte injury in diabetic mice, while podocyte-specific overexpression was protective.","method":"Co-immunoprecipitation, conditional knockout and overexpression mouse models, ubiquitination assays, Western blot","journal":"Metabolism: clinical and experimental","confidence":"High","confidence_rationale":"Tier 1-2 / Moderate — direct interaction demonstrated by Co-IP, ubiquitination mechanism established, validated in both KO and overexpression in vivo models","pmids":["40681143"],"is_preprint":false},{"year":2023,"finding":"Flot2 stabilizes the podocin-nephrin complex by recruiting podocin and nephrin into lipid rafts. Flot2 and podocin directly interact via their SPFH domains. Podocyte-specific Flot2 deletion worsened albuminuria and glomerular pathology in LPS/ADR-induced nephropathy, while podocyte-specific overexpression was protective. Flot2 was identified as a direct transcriptional target of KLF15.","method":"Co-immunoprecipitation, domain mapping (SPFH), conditional knockout and transgenic overexpression mouse models, sucrose density fractionation, ChIP/luciferase for KLF15 regulation","journal":"International journal of biological sciences","confidence":"High","confidence_rationale":"Tier 1-2 / Moderate — direct interaction via SPFH domain, lipid raft fractionation, validated in both KO and transgenic OE models, multiple orthogonal methods","pmids":["36632460"],"is_preprint":false},{"year":2024,"finding":"HDAC6 deacetylates FLOT2 at lysine 211 (K211), inhibiting its proteasomal degradation. Acetylation-mimicking mutation (K211R) slows FLOT2 degradation and increases its tumorigenic activity. HDAC6 and FLOT2 interact (confirmed by Co-IP), and HDAC6 knockdown increases FLOT2 acetylation and reduces FLOT2 protein levels via the proteasome.","method":"Co-immunoprecipitation, site-directed mutagenesis (K211R), cycloheximide chase assay, proteasome inhibitor (MG132) treatment, HDAC inhibitor (TSA) treatment, Western blot","journal":"Journal of Central South University. Medical sciences","confidence":"High","confidence_rationale":"Tier 1 / Moderate — active-site mutagenesis, Co-IP, mechanistic degradation assays with multiple inhibitors, single lab","pmids":["39174882"],"is_preprint":false},{"year":2024,"finding":"HNRNPH1 stabilizes FLOT2 mRNA through an m6A-dependent mechanism: HNRNPH1 interacts with and protects METTL14 from STUB1-mediated degradation, leading to increased m6A modification of FLOT2 mRNA, which is then recognized by IGF2BP3 to further stabilize it. Restoration of METTL14 in HNRNPH1-depleted cells rescues FLOT2 expression and malignant phenotype, but not when FLOT2 is also knocked down.","method":"Co-immunoprecipitation, mRNA stability assays, m6A methylation analysis, rescue experiments with METTL14 and FLOT2, in vitro and in vivo functional assays","journal":"Cellular oncology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — Co-IP, epistatic rescue experiments, m6A modification mechanistic chain, single lab with multiple orthogonal methods","pmids":["39570559"],"is_preprint":false},{"year":2024,"finding":"FTO demethylates m6A modifications in FLOT2 mRNA, stabilizing it and upregulating FLOT2 protein expression, which subsequently activates the PI3K/Akt/mTOR signaling pathway in DLBCL. FTO overexpression promotes and FTO silencing suppresses DLBCL malignant phenotypes through FLOT2.","method":"m6A methylation assays, mRNA stability assays, overexpression/silencing vectors, Western blot, functional assays (viability, apoptosis, invasion), rescue experiments with FLOT2","journal":"Archives of biochemistry and biophysics","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — m6A mechanism established with functional rescue, single lab, multiple methods","pmids":["38914215"],"is_preprint":false},{"year":2017,"finding":"Flot2 promotes HCC tumor growth and metastasis by activating the Raf/MEK/ERK1/2 pathway, upregulating Twist, which drives cell cycle modulation and EMT induction. Forced overexpression of Flot2 promoted proliferation, migration, and invasion, while silencing inhibited these processes both in vitro and in vivo.","method":"Overexpression and siRNA knockdown, Western blot, in vitro functional assays, in vivo xenograft model, pathway analysis","journal":"American journal of cancer research","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — loss- and gain-of-function with pathway readouts, in vivo validation, single lab","pmids":["28560058"],"is_preprint":false},{"year":2019,"finding":"During neural differentiation of P19C6 cells, Flot2 expression and localization to lipid rafts increases. Fyn kinase co-localizes with Flot2 in lipid rafts and partially colocalizes after neural differentiation, suggesting Fyn phosphorylates Flot2 in lipid rafts during neural differentiation.","method":"Sucrose density gradient fractionation, immunofluorescence co-localization, Western blot of detergent-resistant membrane fractions","journal":"BMC molecular and cell biology","confidence":"Medium","confidence_rationale":"Tier 2 / Weak — direct fractionation and co-localization, but phosphorylation by Fyn inferred rather than directly demonstrated; single lab, single study","pmids":["31455216"],"is_preprint":false},{"year":2017,"finding":"LXR agonist T0901317 decreases mRNA and protein expression of FLOT2 and its palmitoylating enzyme DHHC5 in MCF-7 breast cancer cells, and reduces Akt phosphorylation and its localization at the plasma membrane, indicating LXR-mediated transcriptional regulation of FLOT2 affects lipid raft integrity and downstream Akt signaling.","method":"LXR agonist treatment, RT-PCR, Western blot, Akt localization assay","journal":"Anticancer research","confidence":"Low","confidence_rationale":"Tier 3 / Weak — pharmacological treatment with functional readouts, no direct mechanistic dissection of LXR-FLOT2 transcriptional link, single lab, single method per endpoint","pmids":["28739689"],"is_preprint":false},{"year":2022,"finding":"FLOT2 promotes NPC progression by stabilizing STAT3 protein, which then transcriptionally activates CD109 expression. CD109 upregulation suppresses the TGF-β/Smad pathway. Rescue experiments showed CD109 reverses functional changes induced by FLOT2 alteration in vitro and in vivo.","method":"Co-immunoprecipitation, immunofluorescence, ChIP, dual-luciferase assay, qRT-PCR, Western blot, in vitro and in vivo functional assays","journal":"iScience","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — Co-IP, ChIP, and rescue experiments showing mechanistic chain FLOT2→STAT3→CD109→TGF-β, single lab, multiple orthogonal methods","pmids":["38161417"],"is_preprint":false},{"year":2022,"finding":"TBL1X interacts with TCF4 to trans-activate Flot2 expression. Flot2 in turn increases TBL1X expression by upregulating c-myc, a positive transcriptional regulator of TBL1X, forming a positive feedback loop that promotes NPC metastasis.","method":"Co-immunoprecipitation, luciferase reporter assay, qRT-PCR, Western blot, in vitro and in vivo functional assays","journal":"International journal of biological sciences","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — Co-IP and luciferase assay support interaction and transcriptional activation, feedback loop established through rescue experiments, single lab","pmids":["35173544"],"is_preprint":false},{"year":2023,"finding":"Flot2 deficiency in B cells leads to elevated effector B cell (Beff) cytokines (IL-6, IL-1β, CXCL10) without affecting regulatory B cell (Breg) cytokines (IL-10, TGF-β), exacerbating sepsis-induced lung injury and reducing survival. Flot2 acts as a suppressor of Beff inflammatory responses.","method":"B cell-specific Flot2 knockout mice and chimeric mice, RNA-seq, ELISA, in vivo CLP sepsis model","journal":"Immunology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — cell-specific KO with RNA-seq and functional in vivo readouts, single lab","pmids":["37688314"],"is_preprint":false},{"year":2025,"finding":"FLOT2 interacts with PLCγ2 in osteoclast precursor cells; FLOT2 deficiency disrupts this interaction and inhibits osteoclastogenesis. Targeted ablation of Flot2 using CRISPR/Cas9 and siRNA suppressed osteoclastogenesis in vitro and prevented ovariectomy-induced osteoporosis in vivo without affecting physiological bone mass.","method":"CRISPR/Cas9 knockout, siRNA knockdown, Co-immunoprecipitation (FLOT2-PLCγ2), in vitro osteoclastogenesis assays, in vivo OVX mouse model","journal":"Pharmacological research","confidence":"High","confidence_rationale":"Tier 1-2 / Moderate — CRISPR KO, siRNA, Co-IP, and validated in vivo; multiple orthogonal approaches in a single study","pmids":["40816424"],"is_preprint":false},{"year":2015,"finding":"miR-449a directly targets the 3'UTR of Flot2 (validated by luciferase reporter assay), reducing Flot2 expression. Flot2 is necessary for TGF-β-induced EMT in gastric cancer cells; its silencing reduces mesenchymal markers and increases E-cadherin, suppressing invasion.","method":"Luciferase reporter assay, qRT-PCR, Western blot, invasion assays","journal":"Diagnostic pathology","confidence":"Medium","confidence_rationale":"Tier 3 / Moderate — luciferase validated direct targeting, functional epistasis with TGF-β pathway, replicated across multiple cell lines in single lab","pmids":["26576674"],"is_preprint":false},{"year":2023,"finding":"Loss of Flot2 expression in deep cerebellar nuclei neurons of Npc1 knockout mice correlates with lipid raft disruption due to GM2 ganglioside sequestration. Treatment with miglustat (sphingolipid synthesis inhibitor) or N-acetyl-l-leucine restored Flot2 expression in Npc1-deficient mice.","method":"Npc1 knockout mouse model, immunostaining, single-nucleus RNA sequencing, pharmacological treatment","journal":"Heliyon","confidence":"Low","confidence_rationale":"Tier 3 / Weak — descriptive loss of Flot2 in disease model with pharmacological rescue, no direct mechanistic dissection of how Flot2 connects to raft disruption","pmids":["37539272"],"is_preprint":false}],"current_model":"FLOT2 is a lipid raft scaffolding protein that organizes membrane microdomains to regulate multiple signaling pathways: it promotes cytoneme-based Wnt ligand transport (with Ror2), stabilizes receptor/complex partners (EphA2, podocin-nephrin, synaptopodin, PLCγ2) by protecting them from proteasomal degradation, interacts with LAMP2 to facilitate autophagosome-lysosome fusion, and activates downstream oncogenic cascades (Raf/MEK/ERK, PI3K/Akt/mTOR) in multiple cancer contexts; its own stability is regulated post-translationally by HDAC6-mediated deacetylation at K211 (preventing proteasomal degradation) and at the mRNA level by FTO-mediated m6A demethylation and the HNRNPH1/METTL14/IGF2BP3 axis."},"narrative":{"mechanistic_narrative":"FLOT2 is a lipid raft scaffolding protein that organizes membrane microdomains to coordinate intercellular ligand transport, receptor/complex stabilization, and downstream oncogenic and inflammatory signaling [PMID:36040316, PMID:36632460]. At the membrane, it recruits partners into detergent-resistant rafts: it forms a complex with podocin and nephrin through direct SPFH-domain contact to stabilize the slit-diaphragm in podocytes [PMID:36632460], directly binds synaptopodin and protects it from K48-linked polyubiquitination and proteasomal degradation [PMID:40681143], and stabilizes EphA2 to drive Akt/NF-κB signaling in glioma [PMID:33631676]. Acting with the Wnt co-receptor Ror2, FLOT2 promotes cytoneme-based intercellular transport of Wnt ligands, a function conserved from cancer cells to zebrafish embryogenesis [PMID:36040316]. FLOT2 also binds PLCγ2 to support osteoclastogenesis [PMID:40816424] and interacts with LAMP2 to promote autophagosome-lysosome fusion [PMID:40066518]. Across multiple cancers it activates pro-tumorigenic cascades, including Raf/MEK/ERK–Twist-driven EMT [PMID:28560058], PI3K/Akt/mTOR signaling [PMID:38914215], and STAT3-dependent transcriptional programs [PMID:38161417]. FLOT2 abundance is tightly controlled post-translationally—HDAC6 deacetylates FLOT2 at K211 to block its proteasomal degradation [PMID:39174882]—and at the mRNA level through m6A-dependent regulation by the HNRNPH1/METTL14/IGF2BP3 axis [PMID:39570559] and FTO-mediated demethylation [PMID:38914215], as well as by transcriptional inputs and miRNA targeting [PMID:36632460, PMID:35173544, PMID:26576674].","teleology":[{"year":2015,"claim":"Established that FLOT2 is a functionally required driver of EMT and a regulated target, answering whether its expression is rate-limiting for invasion.","evidence":"miR-449a 3'UTR luciferase targeting, silencing, and TGF-β-induced EMT assays in gastric cancer cells","pmids":["26576674"],"confidence":"Medium","gaps":["Does not define the molecular activity by which FLOT2 enables EMT","No in vivo validation in this study"]},{"year":2017,"claim":"Connected FLOT2 to a specific oncogenic kinase cascade, showing how it converts to a proliferative/metastatic phenotype.","evidence":"Gain/loss-of-function with Raf/MEK/ERK1/2 and Twist pathway readouts plus xenograft in HCC; LXR agonist modulation of FLOT2/DHHC5 and Akt in breast cancer cells","pmids":["28560058","28739689"],"confidence":"Medium","gaps":["Direct physical link between FLOT2 and Raf/MEK/ERK components not shown","LXR-FLOT2 transcriptional connection only pharmacologically inferred"]},{"year":2019,"claim":"Linked FLOT2 raft localization to a developmental context, showing dynamic raft partitioning during neural differentiation.","evidence":"Sucrose density fractionation and Fyn co-localization in differentiating P19C6 cells","pmids":["31455216"],"confidence":"Medium","gaps":["Fyn phosphorylation of FLOT2 inferred, not directly demonstrated","Functional consequence of raft enrichment for differentiation untested"]},{"year":2021,"claim":"Demonstrated FLOT2 acts as a receptor-stabilizing scaffold, placing it upstream of EphA2 in a pro-survival axis.","evidence":"Reciprocal Co-IP, siRNA knockdown, and EphA2 rescue in glioma cells","pmids":["33631676"],"confidence":"Medium","gaps":["Mechanism by which FLOT2 protects EphA2 from turnover not defined","Single-lab evidence"]},{"year":2022,"claim":"Defined a membrane-scaffolding role distinct from signal transduction—FLOT2 controls cytoneme-based intercellular Wnt ligand transport with Ror2, conserved in vivo.","evidence":"Live cytoneme imaging, genetic Flot2/Ror2 manipulation, and zebrafish embryogenesis model","pmids":["36040316"],"confidence":"High","gaps":["Biochemical basis of Wnt3 loading onto cytonemes unresolved","Structural requirements of the FLOT2-Ror2 interface not mapped"]},{"year":2022,"claim":"Embedded FLOT2 in transcriptional feedback circuits, showing it both responds to and reinforces oncogenic transcription factor activity.","evidence":"Co-IP, ChIP, luciferase, and rescue experiments delineating FLOT2-STAT3-CD109-TGF-β and TBL1X/TCF4-FLOT2-c-myc loops in NPC","pmids":["38161417","35173544"],"confidence":"Medium","gaps":["How a membrane scaffold stabilizes nuclear/cytoplasmic STAT3 mechanistically unclear","Loops characterized only in NPC contexts"]},{"year":2023,"claim":"Established FLOT2 as a direct lipid-raft organizer of the slit diaphragm via SPFH-domain interaction, with a defined upstream transcriptional regulator.","evidence":"Co-IP, SPFH domain mapping, sucrose fractionation, conditional KO/transgenic mice, and KLF15 ChIP/luciferase","pmids":["36632460"],"confidence":"High","gaps":["Whether raft recruitment alone or additional chaperoning stabilizes the complex unresolved","Stoichiometry of podocin-nephrin-FLOT2 assembly unknown"]},{"year":2023,"claim":"Extended FLOT2 function to immune regulation and to disease-associated raft disruption, showing context-dependent suppressive roles.","evidence":"B cell-specific Flot2 KO with RNA-seq/ELISA in CLP sepsis; descriptive loss of Flot2 in Npc1-null cerebellar neurons with pharmacological rescue","pmids":["37688314","37539272"],"confidence":"Medium","gaps":["Molecular target of FLOT2 in Beff cytokine suppression unidentified","NPC link is correlative without mechanistic dissection"]},{"year":2024,"claim":"Resolved how FLOT2 protein and mRNA abundance are controlled, identifying acetylation- and m6A-based regulatory layers.","evidence":"HDAC6 K211 deacetylation with K211R mutagenesis and CHX/MG132 chase; HNRNPH1/METTL14/IGF2BP3 m6A stabilization and FTO demethylation with rescue assays","pmids":["39174882","39570559","38914215"],"confidence":"Medium","gaps":["E3 ligase mediating FLOT2 proteasomal turnover not identified","Interplay between acetylation and m6A control untested"]},{"year":2025,"claim":"Demonstrated FLOT2 functions as an anti-degradation scaffold in vivo across organ systems and supports receptor-proximal enzyme signaling.","evidence":"Co-IP and conditional KO/OE mice showing synaptopodin protection from K48 ubiquitination in diabetic nephropathy; FLOT2-PLCγ2 Co-IP with CRISPR KO and OVX osteoporosis model; LAMP2-FLOT2 Co-IP and autophagic flux assays in cardiac endothelium","pmids":["40681143","40816424","40066518"],"confidence":"High","gaps":["Whether a single raft-scaffolding mechanism underlies all partner stabilization events unproven","Direct enzymatic or adaptor activity of FLOT2 not established"]},{"year":null,"claim":"It remains unknown whether FLOT2's diverse partner-stabilizing and signaling activities derive from a single unifying biochemical mechanism of raft organization or from distinct context-specific interactions.","evidence":"","pmids":[],"confidence":"Medium","gaps":["No structural model of FLOT2 raft assembly or partner binding","E3 ligases and full degradation machinery acting on FLOT2 and its clients undefined","Direct molecular activity (beyond scaffolding) not demonstrated"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0060090","term_label":"molecular adaptor activity","supporting_discovery_ids":[0,4,1,3,14,2]},{"term_id":"GO:0140313","term_label":"molecular sequestering activity","supporting_discovery_ids":[3,4,1]}],"localization":[{"term_id":"GO:0005886","term_label":"plasma membrane","supporting_discovery_ids":[4,9,10]},{"term_id":"GO:0005856","term_label":"cytoskeleton","supporting_discovery_ids":[0]}],"pathway":[{"term_id":"R-HSA-162582","term_label":"Signal Transduction","supporting_discovery_ids":[1,8,7,11]},{"term_id":"R-HSA-9612973","term_label":"Autophagy","supporting_discovery_ids":[2]},{"term_id":"R-HSA-1643685","term_label":"Disease","supporting_discovery_ids":[3,4,14]}],"complexes":["podocin-nephrin slit diaphragm complex"],"partners":["ROR2","EPHA2","SYNPO","PODOCIN (NPHS2)","NPHS1","LAMP2","PLCG2","HDAC6"],"other_free_text":[]}},"prefetch_data":{"uniprot":{"accession":"Q14254","full_name":"Flotillin-2","aliases":["Epidermal surface antigen","ESA","Membrane component chromosome 17 surface marker 1"],"length_aa":428,"mass_kda":47.1,"function":"May act as a scaffolding protein within caveolar membranes, functionally participating in formation of caveolae or caveolae-like vesicles. May be involved in epidermal cell adhesion and epidermal structure and function","subcellular_location":"Cell membrane; Membrane, caveola; Endosome; Membrane","url":"https://www.uniprot.org/uniprotkb/Q14254/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/FLOT2","classification":"Not Classified","n_dependent_lines":12,"n_total_lines":1208,"dependency_fraction":0.009933774834437087},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[{"gene":"CEP192","stoichiometry":0.2}],"url":"https://opencell.sf.czbiohub.org/search/FLOT2","total_profiled":1310},"omim":[{"mim_id":"614586","title":"ZDHHC PALMITOYLTRANSFERASE 5; ZDHHC5","url":"https://www.omim.org/entry/614586"},{"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":"605973","title":"DDB1- AND CUL4-ASSOCIATED FACTOR 7; DCAF7","url":"https://www.omim.org/entry/605973"},{"mim_id":"131560","title":"FLOTILLIN 2; FLOT2","url":"https://www.omim.org/entry/131560"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"Supported","locations":[{"location":"Plasma membrane","reliability":"Supported"},{"location":"Vesicles","reliability":"Additional"}],"tissue_specificity":"Low tissue specificity","tissue_distribution":"Detected in all","driving_tissues":[],"url":"https://www.proteinatlas.org/search/FLOT2"},"hgnc":{"alias_symbol":["ESA","ESA1","ECS-1","ECS1"],"prev_symbol":["M17S1"]},"alphafold":{"accession":"Q14254","domains":[{"cath_id":"3.30.479.30","chopping":"49-161","consensus_level":"medium","plddt":92.3911,"start":49,"end":161},{"cath_id":"2.30.30","chopping":"1-47","consensus_level":"medium","plddt":83.56,"start":1,"end":47}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/Q14254","model_url":"https://alphafold.ebi.ac.uk/files/AF-Q14254-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-Q14254-F1-predicted_aligned_error_v6.png","plddt_mean":82.0},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=FLOT2","jax_strain_url":"https://www.jax.org/strain/search?query=FLOT2"},"sequence":{"accession":"Q14254","fasta_url":"https://rest.uniprot.org/uniprotkb/Q14254.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/Q14254/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/Q14254"}},"corpus_meta":[{"pmid":"31287224","id":"PMC_31287224","title":"MicroRNA-351 eases insulin resistance and liver gluconeogenesis via the PI3K/AKT pathway by inhibiting FLOT2 in mice of gestational diabetes mellitus.","date":"2019","source":"Journal of cellular and molecular medicine","url":"https://pubmed.ncbi.nlm.nih.gov/31287224","citation_count":69,"is_preprint":false},{"pmid":"27262438","id":"PMC_27262438","title":"MiR-485 inhibits metastasis and EMT of lung adenocarcinoma by targeting Flot2.","date":"2016","source":"Biochemical and biophysical research communications","url":"https://pubmed.ncbi.nlm.nih.gov/27262438","citation_count":48,"is_preprint":false},{"pmid":"26576674","id":"PMC_26576674","title":"miR-449a targets Flot2 and inhibits gastric cancer invasion by inhibiting TGF-β-mediated EMT.","date":"2015","source":"Diagnostic pathology","url":"https://pubmed.ncbi.nlm.nih.gov/26576674","citation_count":39,"is_preprint":false},{"pmid":"25403318","id":"PMC_25403318","title":"Identification of FLOT2 as a novel target for microRNA-34a in 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obesity-associated protein (FTO)-induced upregulation of flotillin-2 (FLOT2) contributes to cancer aggressiveness in diffuse large B-cell lymphoma (DLBCL) via activating the PI3K/Akt/mTOR signal pathway.","date":"2024","source":"Archives of biochemistry and biophysics","url":"https://pubmed.ncbi.nlm.nih.gov/38914215","citation_count":12,"is_preprint":false},{"pmid":"33817236","id":"PMC_33817236","title":"Circ_001569 regulates FLOT2 expression to promote the proliferation, migration, invasion and EMT of osteosarcoma cells through sponging miR-185-5p.","date":"2020","source":"Open life sciences","url":"https://pubmed.ncbi.nlm.nih.gov/33817236","citation_count":12,"is_preprint":false},{"pmid":"31455216","id":"PMC_31455216","title":"The Flot2 component of the lipid raft changes localization during neural differentiation of P19C6 cells.","date":"2019","source":"BMC molecular and cell 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Together with the Wnt co-receptor Ror2, Flot2 determines the number and length of Wnt3-carrying cytonemes. This mechanism is conserved: Flotillins are also necessary for Wnt8a cytonemes during zebrafish embryogenesis.\",\n      \"method\": \"Live imaging of cytonemes, genetic manipulation of Flot2 and Ror2, zebrafish embryogenesis model, functional proliferation/survival assays\",\n      \"journal\": \"eLife\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — multiple orthogonal methods (imaging, genetic manipulation, in vivo zebrafish model), single lab but mechanistically rigorous with functional validation\",\n      \"pmids\": [\"36040316\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"FLOT2 interacts with and stabilizes EphA2 protein in glioma cells; FLOT2 knockdown reduces EphA2 levels, suppresses Akt and NF-κB activities, and induces apoptosis, cell cycle arrest, and EMT inhibition. Restoration of EphA2 rescues the suppressive effects of FLOT2 knockdown, placing FLOT2 upstream of EphA2 in a pro-oncogenic signaling axis.\",\n      \"method\": \"Co-immunoprecipitation, siRNA knockdown, Western blot, IHC, functional rescue assays\",\n      \"journal\": \"Biochemical and biophysical research communications\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — reciprocal Co-IP and functional rescue in single lab with multiple orthogonal methods\",\n      \"pmids\": [\"33631676\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"LAMP2 preferentially binds FLOT2 in cardiac endothelial cells following IL-4 exposure, and this LAMP2-FLOT2 interaction enhances autophagosome-lysosome fusion. Loss of FLOT2 reverses the LAMP2-mediated rescue of autophagic flux, leading to autophagosome accumulation.\",\n      \"method\": \"Co-immunoprecipitation, loss-of-function (FLOT2 knockdown), autophagic flux assays (including transmission electron microscopy and LC3/p62 markers), CLP mouse sepsis model\",\n      \"journal\": \"Autophagy\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — Co-IP, functional KO, multiple orthogonal readouts (TEM, flux markers), single lab\",\n      \"pmids\": [\"40066518\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"Flot2 directly interacts with synaptopodin in podocytes and protects it from proteasomal degradation via inhibition of K48-linked polyubiquitination. Podocyte-specific Flot2 knockout worsened albuminuria and podocyte injury in diabetic mice, while podocyte-specific overexpression was protective.\",\n      \"method\": \"Co-immunoprecipitation, conditional knockout and overexpression mouse models, ubiquitination assays, Western blot\",\n      \"journal\": \"Metabolism: clinical and experimental\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1-2 / Moderate — direct interaction demonstrated by Co-IP, ubiquitination mechanism established, validated in both KO and overexpression in vivo models\",\n      \"pmids\": [\"40681143\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"Flot2 stabilizes the podocin-nephrin complex by recruiting podocin and nephrin into lipid rafts. Flot2 and podocin directly interact via their SPFH domains. Podocyte-specific Flot2 deletion worsened albuminuria and glomerular pathology in LPS/ADR-induced nephropathy, while podocyte-specific overexpression was protective. Flot2 was identified as a direct transcriptional target of KLF15.\",\n      \"method\": \"Co-immunoprecipitation, domain mapping (SPFH), conditional knockout and transgenic overexpression mouse models, sucrose density fractionation, ChIP/luciferase for KLF15 regulation\",\n      \"journal\": \"International journal of biological sciences\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1-2 / Moderate — direct interaction via SPFH domain, lipid raft fractionation, validated in both KO and transgenic OE models, multiple orthogonal methods\",\n      \"pmids\": [\"36632460\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"HDAC6 deacetylates FLOT2 at lysine 211 (K211), inhibiting its proteasomal degradation. Acetylation-mimicking mutation (K211R) slows FLOT2 degradation and increases its tumorigenic activity. HDAC6 and FLOT2 interact (confirmed by Co-IP), and HDAC6 knockdown increases FLOT2 acetylation and reduces FLOT2 protein levels via the proteasome.\",\n      \"method\": \"Co-immunoprecipitation, site-directed mutagenesis (K211R), cycloheximide chase assay, proteasome inhibitor (MG132) treatment, HDAC inhibitor (TSA) treatment, Western blot\",\n      \"journal\": \"Journal of Central South University. Medical sciences\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — active-site mutagenesis, Co-IP, mechanistic degradation assays with multiple inhibitors, single lab\",\n      \"pmids\": [\"39174882\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"HNRNPH1 stabilizes FLOT2 mRNA through an m6A-dependent mechanism: HNRNPH1 interacts with and protects METTL14 from STUB1-mediated degradation, leading to increased m6A modification of FLOT2 mRNA, which is then recognized by IGF2BP3 to further stabilize it. Restoration of METTL14 in HNRNPH1-depleted cells rescues FLOT2 expression and malignant phenotype, but not when FLOT2 is also knocked down.\",\n      \"method\": \"Co-immunoprecipitation, mRNA stability assays, m6A methylation analysis, rescue experiments with METTL14 and FLOT2, in vitro and in vivo functional assays\",\n      \"journal\": \"Cellular oncology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — Co-IP, epistatic rescue experiments, m6A modification mechanistic chain, single lab with multiple orthogonal methods\",\n      \"pmids\": [\"39570559\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"FTO demethylates m6A modifications in FLOT2 mRNA, stabilizing it and upregulating FLOT2 protein expression, which subsequently activates the PI3K/Akt/mTOR signaling pathway in DLBCL. FTO overexpression promotes and FTO silencing suppresses DLBCL malignant phenotypes through FLOT2.\",\n      \"method\": \"m6A methylation assays, mRNA stability assays, overexpression/silencing vectors, Western blot, functional assays (viability, apoptosis, invasion), rescue experiments with FLOT2\",\n      \"journal\": \"Archives of biochemistry and biophysics\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — m6A mechanism established with functional rescue, single lab, multiple methods\",\n      \"pmids\": [\"38914215\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"Flot2 promotes HCC tumor growth and metastasis by activating the Raf/MEK/ERK1/2 pathway, upregulating Twist, which drives cell cycle modulation and EMT induction. Forced overexpression of Flot2 promoted proliferation, migration, and invasion, while silencing inhibited these processes both in vitro and in vivo.\",\n      \"method\": \"Overexpression and siRNA knockdown, Western blot, in vitro functional assays, in vivo xenograft model, pathway analysis\",\n      \"journal\": \"American journal of cancer research\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — loss- and gain-of-function with pathway readouts, in vivo validation, single lab\",\n      \"pmids\": [\"28560058\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"During neural differentiation of P19C6 cells, Flot2 expression and localization to lipid rafts increases. Fyn kinase co-localizes with Flot2 in lipid rafts and partially colocalizes after neural differentiation, suggesting Fyn phosphorylates Flot2 in lipid rafts during neural differentiation.\",\n      \"method\": \"Sucrose density gradient fractionation, immunofluorescence co-localization, Western blot of detergent-resistant membrane fractions\",\n      \"journal\": \"BMC molecular and cell biology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Weak — direct fractionation and co-localization, but phosphorylation by Fyn inferred rather than directly demonstrated; single lab, single study\",\n      \"pmids\": [\"31455216\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"LXR agonist T0901317 decreases mRNA and protein expression of FLOT2 and its palmitoylating enzyme DHHC5 in MCF-7 breast cancer cells, and reduces Akt phosphorylation and its localization at the plasma membrane, indicating LXR-mediated transcriptional regulation of FLOT2 affects lipid raft integrity and downstream Akt signaling.\",\n      \"method\": \"LXR agonist treatment, RT-PCR, Western blot, Akt localization assay\",\n      \"journal\": \"Anticancer research\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — pharmacological treatment with functional readouts, no direct mechanistic dissection of LXR-FLOT2 transcriptional link, single lab, single method per endpoint\",\n      \"pmids\": [\"28739689\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"FLOT2 promotes NPC progression by stabilizing STAT3 protein, which then transcriptionally activates CD109 expression. CD109 upregulation suppresses the TGF-β/Smad pathway. Rescue experiments showed CD109 reverses functional changes induced by FLOT2 alteration in vitro and in vivo.\",\n      \"method\": \"Co-immunoprecipitation, immunofluorescence, ChIP, dual-luciferase assay, qRT-PCR, Western blot, in vitro and in vivo functional assays\",\n      \"journal\": \"iScience\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — Co-IP, ChIP, and rescue experiments showing mechanistic chain FLOT2→STAT3→CD109→TGF-β, single lab, multiple orthogonal methods\",\n      \"pmids\": [\"38161417\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"TBL1X interacts with TCF4 to trans-activate Flot2 expression. Flot2 in turn increases TBL1X expression by upregulating c-myc, a positive transcriptional regulator of TBL1X, forming a positive feedback loop that promotes NPC metastasis.\",\n      \"method\": \"Co-immunoprecipitation, luciferase reporter assay, qRT-PCR, Western blot, in vitro and in vivo functional assays\",\n      \"journal\": \"International journal of biological sciences\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — Co-IP and luciferase assay support interaction and transcriptional activation, feedback loop established through rescue experiments, single lab\",\n      \"pmids\": [\"35173544\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"Flot2 deficiency in B cells leads to elevated effector B cell (Beff) cytokines (IL-6, IL-1β, CXCL10) without affecting regulatory B cell (Breg) cytokines (IL-10, TGF-β), exacerbating sepsis-induced lung injury and reducing survival. Flot2 acts as a suppressor of Beff inflammatory responses.\",\n      \"method\": \"B cell-specific Flot2 knockout mice and chimeric mice, RNA-seq, ELISA, in vivo CLP sepsis model\",\n      \"journal\": \"Immunology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — cell-specific KO with RNA-seq and functional in vivo readouts, single lab\",\n      \"pmids\": [\"37688314\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"FLOT2 interacts with PLCγ2 in osteoclast precursor cells; FLOT2 deficiency disrupts this interaction and inhibits osteoclastogenesis. Targeted ablation of Flot2 using CRISPR/Cas9 and siRNA suppressed osteoclastogenesis in vitro and prevented ovariectomy-induced osteoporosis in vivo without affecting physiological bone mass.\",\n      \"method\": \"CRISPR/Cas9 knockout, siRNA knockdown, Co-immunoprecipitation (FLOT2-PLCγ2), in vitro osteoclastogenesis assays, in vivo OVX mouse model\",\n      \"journal\": \"Pharmacological research\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1-2 / Moderate — CRISPR KO, siRNA, Co-IP, and validated in vivo; multiple orthogonal approaches in a single study\",\n      \"pmids\": [\"40816424\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2015,\n      \"finding\": \"miR-449a directly targets the 3'UTR of Flot2 (validated by luciferase reporter assay), reducing Flot2 expression. Flot2 is necessary for TGF-β-induced EMT in gastric cancer cells; its silencing reduces mesenchymal markers and increases E-cadherin, suppressing invasion.\",\n      \"method\": \"Luciferase reporter assay, qRT-PCR, Western blot, invasion assays\",\n      \"journal\": \"Diagnostic pathology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 / Moderate — luciferase validated direct targeting, functional epistasis with TGF-β pathway, replicated across multiple cell lines in single lab\",\n      \"pmids\": [\"26576674\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"Loss of Flot2 expression in deep cerebellar nuclei neurons of Npc1 knockout mice correlates with lipid raft disruption due to GM2 ganglioside sequestration. Treatment with miglustat (sphingolipid synthesis inhibitor) or N-acetyl-l-leucine restored Flot2 expression in Npc1-deficient mice.\",\n      \"method\": \"Npc1 knockout mouse model, immunostaining, single-nucleus RNA sequencing, pharmacological treatment\",\n      \"journal\": \"Heliyon\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — descriptive loss of Flot2 in disease model with pharmacological rescue, no direct mechanistic dissection of how Flot2 connects to raft disruption\",\n      \"pmids\": [\"37539272\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"FLOT2 is a lipid raft scaffolding protein that organizes membrane microdomains to regulate multiple signaling pathways: it promotes cytoneme-based Wnt ligand transport (with Ror2), stabilizes receptor/complex partners (EphA2, podocin-nephrin, synaptopodin, PLCγ2) by protecting them from proteasomal degradation, interacts with LAMP2 to facilitate autophagosome-lysosome fusion, and activates downstream oncogenic cascades (Raf/MEK/ERK, PI3K/Akt/mTOR) in multiple cancer contexts; its own stability is regulated post-translationally by HDAC6-mediated deacetylation at K211 (preventing proteasomal degradation) and at the mRNA level by FTO-mediated m6A demethylation and the HNRNPH1/METTL14/IGF2BP3 axis.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"FLOT2 is a lipid raft scaffolding protein that organizes membrane microdomains to coordinate intercellular ligand transport, receptor/complex stabilization, and downstream oncogenic and inflammatory signaling [#0, #4]. At the membrane, it recruits partners into detergent-resistant rafts: it forms a complex with podocin and nephrin through direct SPFH-domain contact to stabilize the slit-diaphragm in podocytes [#4], directly binds synaptopodin and protects it from K48-linked polyubiquitination and proteasomal degradation [#3], and stabilizes EphA2 to drive Akt/NF-\\u03baB signaling in glioma [#1]. Acting with the Wnt co-receptor Ror2, FLOT2 promotes cytoneme-based intercellular transport of Wnt ligands, a function conserved from cancer cells to zebrafish embryogenesis [#0]. FLOT2 also binds PLC\\u03b32 to support osteoclastogenesis [#14] and interacts with LAMP2 to promote autophagosome-lysosome fusion [#2]. Across multiple cancers it activates pro-tumorigenic cascades, including Raf/MEK/ERK\\u2013Twist-driven EMT [#8], PI3K/Akt/mTOR signaling [#7], and STAT3-dependent transcriptional programs [#11]. FLOT2 abundance is tightly controlled post-translationally\\u2014HDAC6 deacetylates FLOT2 at K211 to block its proteasomal degradation [#5]\\u2014and at the mRNA level through m6A-dependent regulation by the HNRNPH1/METTL14/IGF2BP3 axis [#6] and FTO-mediated demethylation [#7], as well as by transcriptional inputs and miRNA targeting [#4, #12, #15].\",\n  \"teleology\": [\n    {\n      \"year\": 2015,\n      \"claim\": \"Established that FLOT2 is a functionally required driver of EMT and a regulated target, answering whether its expression is rate-limiting for invasion.\",\n      \"evidence\": \"miR-449a 3'UTR luciferase targeting, silencing, and TGF-\\u03b2-induced EMT assays in gastric cancer cells\",\n      \"pmids\": [\"26576674\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Does not define the molecular activity by which FLOT2 enables EMT\", \"No in vivo validation in this study\"]\n    },\n    {\n      \"year\": 2017,\n      \"claim\": \"Connected FLOT2 to a specific oncogenic kinase cascade, showing how it converts to a proliferative/metastatic phenotype.\",\n      \"evidence\": \"Gain/loss-of-function with Raf/MEK/ERK1/2 and Twist pathway readouts plus xenograft in HCC; LXR agonist modulation of FLOT2/DHHC5 and Akt in breast cancer cells\",\n      \"pmids\": [\"28560058\", \"28739689\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Direct physical link between FLOT2 and Raf/MEK/ERK components not shown\", \"LXR-FLOT2 transcriptional connection only pharmacologically inferred\"]\n    },\n    {\n      \"year\": 2019,\n      \"claim\": \"Linked FLOT2 raft localization to a developmental context, showing dynamic raft partitioning during neural differentiation.\",\n      \"evidence\": \"Sucrose density fractionation and Fyn co-localization in differentiating P19C6 cells\",\n      \"pmids\": [\"31455216\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Fyn phosphorylation of FLOT2 inferred, not directly demonstrated\", \"Functional consequence of raft enrichment for differentiation untested\"]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"Demonstrated FLOT2 acts as a receptor-stabilizing scaffold, placing it upstream of EphA2 in a pro-survival axis.\",\n      \"evidence\": \"Reciprocal Co-IP, siRNA knockdown, and EphA2 rescue in glioma cells\",\n      \"pmids\": [\"33631676\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Mechanism by which FLOT2 protects EphA2 from turnover not defined\", \"Single-lab evidence\"]\n    },\n    {\n      \"year\": 2022,\n      \"claim\": \"Defined a membrane-scaffolding role distinct from signal transduction\\u2014FLOT2 controls cytoneme-based intercellular Wnt ligand transport with Ror2, conserved in vivo.\",\n      \"evidence\": \"Live cytoneme imaging, genetic Flot2/Ror2 manipulation, and zebrafish embryogenesis model\",\n      \"pmids\": [\"36040316\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Biochemical basis of Wnt3 loading onto cytonemes unresolved\", \"Structural requirements of the FLOT2-Ror2 interface not mapped\"]\n    },\n    {\n      \"year\": 2022,\n      \"claim\": \"Embedded FLOT2 in transcriptional feedback circuits, showing it both responds to and reinforces oncogenic transcription factor activity.\",\n      \"evidence\": \"Co-IP, ChIP, luciferase, and rescue experiments delineating FLOT2-STAT3-CD109-TGF-\\u03b2 and TBL1X/TCF4-FLOT2-c-myc loops in NPC\",\n      \"pmids\": [\"38161417\", \"35173544\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"How a membrane scaffold stabilizes nuclear/cytoplasmic STAT3 mechanistically unclear\", \"Loops characterized only in NPC contexts\"]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Established FLOT2 as a direct lipid-raft organizer of the slit diaphragm via SPFH-domain interaction, with a defined upstream transcriptional regulator.\",\n      \"evidence\": \"Co-IP, SPFH domain mapping, sucrose fractionation, conditional KO/transgenic mice, and KLF15 ChIP/luciferase\",\n      \"pmids\": [\"36632460\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether raft recruitment alone or additional chaperoning stabilizes the complex unresolved\", \"Stoichiometry of podocin-nephrin-FLOT2 assembly unknown\"]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Extended FLOT2 function to immune regulation and to disease-associated raft disruption, showing context-dependent suppressive roles.\",\n      \"evidence\": \"B cell-specific Flot2 KO with RNA-seq/ELISA in CLP sepsis; descriptive loss of Flot2 in Npc1-null cerebellar neurons with pharmacological rescue\",\n      \"pmids\": [\"37688314\", \"37539272\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Molecular target of FLOT2 in Beff cytokine suppression unidentified\", \"NPC link is correlative without mechanistic dissection\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Resolved how FLOT2 protein and mRNA abundance are controlled, identifying acetylation- and m6A-based regulatory layers.\",\n      \"evidence\": \"HDAC6 K211 deacetylation with K211R mutagenesis and CHX/MG132 chase; HNRNPH1/METTL14/IGF2BP3 m6A stabilization and FTO demethylation with rescue assays\",\n      \"pmids\": [\"39174882\", \"39570559\", \"38914215\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"E3 ligase mediating FLOT2 proteasomal turnover not identified\", \"Interplay between acetylation and m6A control untested\"]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Demonstrated FLOT2 functions as an anti-degradation scaffold in vivo across organ systems and supports receptor-proximal enzyme signaling.\",\n      \"evidence\": \"Co-IP and conditional KO/OE mice showing synaptopodin protection from K48 ubiquitination in diabetic nephropathy; FLOT2-PLC\\u03b32 Co-IP with CRISPR KO and OVX osteoporosis model; LAMP2-FLOT2 Co-IP and autophagic flux assays in cardiac endothelium\",\n      \"pmids\": [\"40681143\", \"40816424\", \"40066518\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether a single raft-scaffolding mechanism underlies all partner stabilization events unproven\", \"Direct enzymatic or adaptor activity of FLOT2 not established\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"It remains unknown whether FLOT2's diverse partner-stabilizing and signaling activities derive from a single unifying biochemical mechanism of raft organization or from distinct context-specific interactions.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No structural model of FLOT2 raft assembly or partner binding\", \"E3 ligases and full degradation machinery acting on FLOT2 and its clients undefined\", \"Direct molecular activity (beyond scaffolding) not demonstrated\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0060090\", \"supporting_discovery_ids\": [0, 4, 1, 3, 14, 2]},\n      {\"term_id\": \"GO:0140313\", \"supporting_discovery_ids\": [3, 4, 1]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005886\", \"supporting_discovery_ids\": [4, 9, 10]},\n      {\"term_id\": \"GO:0005856\", \"supporting_discovery_ids\": [0]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-162582\", \"supporting_discovery_ids\": [1, 8, 7, 11]},\n      {\"term_id\": \"R-HSA-9612973\", \"supporting_discovery_ids\": [2]},\n      {\"term_id\": \"R-HSA-1643685\", \"supporting_discovery_ids\": [3, 4, 14]}\n    ],\n    \"complexes\": [\"podocin-nephrin slit diaphragm complex\"],\n    \"partners\": [\"ROR2\", \"EPHA2\", \"SYNPO\", \"PODOCIN (NPHS2)\", \"NPHS1\", \"LAMP2\", \"PLCG2\", \"HDAC6\"],\n    \"other_free_text\": []\n  }\n}","audit_flag":null,"evaluation":{"pairwise":"win","faith_supported":5,"faith_total":6,"faith_pct":83.33333333333333}}