{"gene":"AFAP1","run_date":"2026-06-09T22:02:42","timeline":{"discoveries":[{"year":2004,"finding":"AFAP-110 (AFAP1) is required to mediate PKCα activation of c-Src and subsequent podosome formation. PKCα activation directs AFAP-110 to colocalize with and bind the c-Src SH3 domain, activating the tyrosine kinase. In cells lacking AFAP-110 (CaOV3), PMA could not activate c-Src or induce podosomes; ectopic AFAP-110 rescued these effects. AFAP-110 mutants unable to bind or colocalize with c-Src failed to rescue.","method":"Immunofluorescence, ectopic expression of constitutively active PKCα, rescue experiments with AFAP-110 mutants in AFAP-110-deficient CaOV3 cells","journal":"Molecular and cellular biology","confidence":"High","confidence_rationale":"Tier 2 / Strong — clean loss-of-function cell line + rescue with multiple domain mutants + defined cellular phenotype, single rigorous study","pmids":["15314167"],"is_preprint":false},{"year":2001,"finding":"AFAP-110 functions as an adaptor protein linking Src family kinases to actin filaments via: (1) SH3/SH2 binding modules in its amino terminus, (2) two pleckstrin homology domains, (3) a leucine zipper motif mediating self-association, (4) a carboxy-terminal actin-binding domain enabling direct actin filament interaction. Conformational changes associated with phosphorylation or mutagenesis reveal an intrinsic capability to alter actin filament integrity and activate cSrc.","method":"Review of prior biochemical, mutagenesis, and cell biology experiments including co-IP, pulldown, in vitro actin binding, and mutational analysis","journal":"Oncogene","confidence":"High","confidence_rationale":"Tier 2 / Strong — synthesis of multiple orthogonal experimental approaches (pulldown, mutagenesis, in vitro binding, immunofluorescence) replicated across multiple papers","pmids":["11607843"],"is_preprint":false},{"year":1997,"finding":"The amino-terminal SH3 binding motif I (SH3bm I) of AFAP-110 is required for stable complex formation with Src and for efficient tyrosine phosphorylation by Src. A proline-to-alanine mutation in SH3bm I abolished GST-SH3 absorption and reduced tyrosine phosphorylation ~2.7-fold, and prevented stable complex formation with activated Src (pp60-527F).","method":"Site-directed mutagenesis of SH3 binding motif, GST-SH3 pulldown, co-expression of Src527F with AFAP-110 mutants in Cos-1 cells, co-immunoprecipitation","journal":"Molecular and cellular biochemistry","confidence":"High","confidence_rationale":"Tier 1 / Moderate — in vitro pulldown + mutagenesis + co-IP, single lab with multiple orthogonal methods","pmids":["9350057"],"is_preprint":false},{"year":1998,"finding":"The carboxy-terminal 127 amino acids of AFAP-110 contain an alpha-helical leucine zipper motif required for self-association (multimerization). Activated Src (Src527F) disrupts carboxy-terminal interactions, shifting AFAP-110 to a multimeric population. Deletion of the leucine zipper region repositions actin filaments into rosette-like structures similar to Src-transformed cells and promotes lamellipodia-like structures. The carboxy terminus is required for colocalization with actin filaments and the cell membrane.","method":"GST fusion protein affinity absorption, Superose size-exclusion chromatography, deletional mutagenesis, immunofluorescence in cells co-expressing Src527F","journal":"Oncogene","confidence":"High","confidence_rationale":"Tier 1 / Moderate — in vitro pulldown reconstitution + size-exclusion chromatography + mutagenesis + cellular imaging, single lab multiple methods","pmids":["9619827"],"is_preprint":false},{"year":2000,"finding":"The carboxy terminus of AFAP-110 is both necessary and sufficient for direct actin filament association in vivo and in vitro. The carboxy-terminal actin-binding domain shows ~40% similarity to known actin-binding motifs. Deletion of the leucine zipper motif (AFAP-Δlzip) enables AFAP-110 to alter actin filament integrity and induce lamellipodia formation in untransformed cells. The leucine zipper motif normally constrains these activities.","method":"In vitro actin filament binding assay with recombinant carboxy-terminal domain, deletional mutagenesis, cellular expression with immunofluorescence in untransformed fibroblasts","journal":"Experimental cell research","confidence":"High","confidence_rationale":"Tier 1 / Moderate — in vitro actin binding reconstitution + mutagenesis + cellular phenotype, single lab with multiple orthogonal methods","pmids":["10666339"],"is_preprint":false},{"year":2001,"finding":"AFAP-110(Δlzip), a leucine zipper deletion mutant, activates cellular tyrosine kinases including Src family members and becomes hyperphosphorylated on tyrosine. This activation requires integrity of the SH3-binding motif of AFAP-110(Δlzip). A deletion within the PH domain also reverts the effects on actin filaments. Dominant-positive RhoA(V14) blocks AFAP-110(Δlzip)-induced actin filament rosettes but not Src activation, placing Rho downstream of AFAP-110-directed Src activation in the actin reorganization pathway.","method":"Expression of deletion and point mutants, immunofluorescence, kinase activity assays, co-expression with dominant-active RhoA","journal":"Oncogene","confidence":"High","confidence_rationale":"Tier 2 / Moderate — epistasis via dominant-active RhoA + mutagenesis + cellular phenotype, single lab multiple methods","pmids":["11641786"],"is_preprint":false},{"year":2002,"finding":"Recombinant AFAP-110 binds actin filaments cooperatively through lateral association and can cross-link actin filaments in a manner dependent on the carboxy-terminal actin-binding domain. Deletion of the leucine zipper or PKC phosphorylation alters AFAP-110 conformation, reduces multimerization stability, and increases its capacity to cross-link actin filaments. PKC activation in cells causes loss of stress fibers, formation of motility structures, and strong AFAP-110 colocalization with these structures. Expression of an AFAP-110 mutant unable to bind PKC blocked PMA effects on actin filaments.","method":"In vitro actin cross-linking assay with recombinant AFAP-110, PKC phosphorylation in vitro, Superose chromatography, expression of PKC-binding domain deletion mutant, phorbol ester treatment and immunofluorescence","journal":"Molecular biology of the cell","confidence":"High","confidence_rationale":"Tier 1 / Moderate — in vitro reconstitution of actin cross-linking + phosphorylation + mutagenesis + cellular imaging, single lab, multiple orthogonal methods","pmids":["12134071"],"is_preprint":false},{"year":1998,"finding":"AFAP-110 forms a stable complex with activated Src through either of two independently functional SH2-binding motifs: one in the amino terminus (Y94) and one in the carboxy terminus (Y451 and Y453). Mutation of Y93/Y94 or Y451/Y453 alone does not fully abrogate complex formation, but triple-tyrosine mutations (combining both regions) drastically reduce phosphorylation and prevent stable complex formation. A multistep SH2/SH3 binding mechanism is supported.","method":"Site-directed mutagenesis of candidate SH2-binding tyrosines, co-expression with Src527F in Cos-1 cells, co-immunoprecipitation, tyrosine phosphorylation analysis","journal":"Molecular carcinogenesis","confidence":"High","confidence_rationale":"Tier 1 / Moderate — systematic site-directed mutagenesis combined with co-IP and phosphorylation assays, single lab multiple complementary approaches","pmids":["9655255"],"is_preprint":false},{"year":2004,"finding":"The leucine zipper motif of AFAP-110 regulates actin filament cross-linking and Src activation through both sequence and structural requirements. The leucine zipper facilitates intramolecular auto-inhibition by contacting the amino-terminal PH1 domain. Disruption of multimer stability (by Δlzip, fos-Lzip substitution, or L581P helix mutation) increases in vitro actin cross-linking; however, only structural disruptions (Δlzip, L581P) activate cSrc and alter actin integrity in vivo, indicating that actin cross-linking and Src activation are distinct functions.","method":"Leucine zipper substitution and point mutagenesis, in vitro actin cross-linking assay, cellular expression with immunofluorescence and cSrc activation assays, opposing binding site analysis","journal":"Journal of cellular biochemistry","confidence":"High","confidence_rationale":"Tier 1 / Moderate — in vitro reconstitution + multiple mutagenesis strategies + cellular phenotype, single lab with clear mechanistic dissection","pmids":["14755689"],"is_preprint":false},{"year":2007,"finding":"AFAP-110 (AFAP1) is required for actin stress fiber formation and cell adhesion to fibronectin in MDA-MB-231 breast cancer cells. Knockdown of AFAP-110 abolishes stress fiber cross-linking and focal adhesion formation without affecting proliferation or expression of focal adhesion components. LPA treatment cannot rescue stress fiber and focal adhesion formation in AFAP-110 knockdown cells.","method":"siRNA knockdown, inducible knockdown, immunofluorescence, fibronectin adhesion assay, LPA stimulation","journal":"Journal of cellular physiology","confidence":"High","confidence_rationale":"Tier 2 / Moderate — clean RNAi knockdown with inducible confirmation + defined cellular phenotype (stress fibers, focal adhesions), single lab two orthogonal methods","pmids":["17520695"],"is_preprint":false},{"year":2007,"finding":"AFAP-110 (AFAP1) is required for tumorigenic growth of prostate cancer cells, regulating focal contacts in a PKC-dependent manner. Downregulation of AFAP-110 reduced cell-matrix adhesion, migration, defective focal adhesions, and reduced integrin β1 expression. Reintroduction of avian AFAP-110 or a Src-interaction-disabled mutant restored these properties, but an AFAP-110 lacking the PKC-interacting domain failed to restore them, demonstrating PKC-dependent regulation of focal contacts.","method":"Immunohistochemistry on human tissue arrays, siRNA knockdown, orthotopic nude mouse model, reintroduction of AFAP-110 domain mutants, cell adhesion and migration assays, integrin β1 expression analysis","journal":"The Journal of clinical investigation","confidence":"High","confidence_rationale":"Tier 2 / Strong — domain mutant rescue experiments + in vivo orthotopic model + multiple phenotypic readouts, single rigorous study","pmids":["17885682"],"is_preprint":false},{"year":2007,"finding":"PI3K activation is required for PMA/PKCα-induced colocalization of AFAP-110 with c-Src and subsequent c-Src activation. The amino-terminal PH1 domain of AFAP-110 is required for colocalization with c-Src, while the SH3-binding motif is required for activation. PI3K inhibitors and cells lacking p85α/β PI3K regulatory subunits block AFAP-110–c-Src colocalization and activation. This pathway promotes cell migration.","method":"PI3K inhibitor treatment, PI3K p85 knockout cells, dominant-negative AFAP-110 expression, wound healing migration assay, immunofluorescence, domain mutant analysis","journal":"American journal of physiology. Cell physiology","confidence":"High","confidence_rationale":"Tier 2 / Moderate — genetic (knockout) and pharmacological PI3K inhibition + domain mutant rescue + migration assay, single lab multiple orthogonal approaches","pmids":["17360811"],"is_preprint":false},{"year":2008,"finding":"PKCα phosphorylates AFAP-110 at Ser277 in response to phorbol ester treatment, and phospho-Ser277 AFAP-110 localizes to podosomes. Mutation of Ser277 to Ala (AFAP-110-S277A) does not alter podosome localization but increases podosome number and lifespan (demonstrated by video microscopy), indicating that Ser277 phosphorylation regulates podosome stability and turnover.","method":"Mutational analysis, phospho-specific antibody generation and immunolabeling, video microscopy of podosome dynamics in A7r5 vascular smooth muscle cells","journal":"Journal of cell science","confidence":"High","confidence_rationale":"Tier 2 / Moderate — phospho-specific antibody validation + mutagenesis + live-cell video microscopy, single lab multiple methods","pmids":["18577577"],"is_preprint":false},{"year":2009,"finding":"Tks5 recruits AFAP-110, p190RhoGAP, and cortactin to podosomes via its fifth SH3 domain. Mislocalization of Tks5 to mitochondria causes redistribution of AFAP-110, p190RhoGAP, and cortactin to perinuclear regions and inhibits podosome formation. A Tks5 mutant lacking the PX domain also inhibits podosome formation and redistributes AFAP-110.","method":"Forced mitochondrial mislocalization of Tks5 deletion mutants, immunofluorescence, siRNA knockdown of Tks5 and p190RhoGAP","journal":"Experimental cell research","confidence":"High","confidence_rationale":"Tier 2 / Moderate — subcellular mislocalization experiment + domain deletion mutant series + siRNA knockdown, single lab multiple approaches","pmids":["19540230"],"is_preprint":false},{"year":1995,"finding":"An alternatively spliced brain-specific variant of AFAP-110, designated AFAP-120, contains an additional 258 bp open reading frame encoding a novel internal sequence (NINS) with a proline-rich SH3-binding motif. The NINS-GST fusion protein pulls down a 67-kDa protein from tissue lysates; deletion analysis maps this interaction to the proline-rich motif.","method":"cDNA cloning, northern blot, western blot, GST-NINS pulldown assay with deletion mutants, transient expression","journal":"The Journal of biological chemistry","confidence":"Medium","confidence_rationale":"Tier 3 / Moderate — single pulldown experiment identifying novel binding partner, corroborated by deletion analysis, single lab","pmids":["7876134"],"is_preprint":false},{"year":2011,"finding":"Endogenous AFAP1 localizes to both cytoskeletal and cytosolic fractions; depolymerization of actin filaments (cytochalasin D or latrunculin A) increases AFAP1 in the cytosolic fraction, demonstrating actin-dependent localization. An AFAP1 mutant lacking the actin-binding domain (AFAPΔABD) localizes predominantly to the cytosolic fraction, is associated with disorganized cytoskeleton, reduced stress fibers, and accumulation of F-actin at the membrane and podosome-like structures that co-localize with cortactin but do not digest gelatin, and show reduced interaction with c-Src.","method":"Subcellular fractionation, cytoskeletal drug treatment, overexpression of ABD deletion mutant, immunofluorescence, gelatin degradation assay","journal":"Cellular and molecular life sciences : CMLS","confidence":"High","confidence_rationale":"Tier 2 / Moderate — fractionation with functional drug perturbations + mutagenesis + multiple cellular phenotype readouts, single lab with multiple orthogonal methods","pmids":["21984596"],"is_preprint":false},{"year":2011,"finding":"AFAP1L1, a paralog of AFAP1 in the AFAP family, preferentially interacts with the SH3 domain of cortactin (not c-Src), in contrast to AFAP1 which binds cSrc SH3 but not cortactin. Overexpression of AFAP1L1 in A7r5 cells induces podosome formation without stimulation and colocalizes with cortactin at invadosomes.","method":"SH3 domain binding assay, fluorescence microscopy, overexpression in A7r5 cells, immunohistochemistry","journal":"European journal of cell biology","confidence":"Medium","confidence_rationale":"Tier 3 / Moderate — SH3 binding specificity comparison + cellular overexpression phenotype; this is primarily about AFAP1L1 but establishes AFAP1's selectivity for cSrc SH3 over cortactin by direct comparison","pmids":["21333378"],"is_preprint":false},{"year":2017,"finding":"AFAP1 (the protein) is a key mediator of TNF-α-induced rapid attenuation of P-glycoprotein (P-gp) efflux activity at the blood-brain barrier. TNF-α rapidly decreases P-gp efflux activity without changing P-gp protein expression. Phosphoproteomics identified AFAP1 as significantly phosphorylated upon TNF-α treatment. Knockdown of AFAP1 blocked the TNF-α-induced reduction in P-gp efflux activity, while MAPK1 inhibition or AP-1 knockdown did not, demonstrating a specific non-transcriptional role for AFAP1 in inflammatory modulation of P-gp function.","method":"Quantitative phosphoproteomics, siRNA knockdown of AFAP1, P-gp efflux activity assay, western blot for P-gp expression in plasma membrane fraction, in vivo LPS mouse model","journal":"Journal of neurochemistry","confidence":"High","confidence_rationale":"Tier 2 / Strong — quantitative phosphoproteomics + selective siRNA knockdown with functional readout + in vivo validation + multiple negative controls (MAPK1, AP-1), single rigorous study","pmids":["28112407"],"is_preprint":false}],"current_model":"AFAP1 (AFAP-110) is an adaptor and actin cross-linking protein that integrates PKCα and PI3K signals to activate c-Src and reorganize the actin cytoskeleton: PKCα phosphorylates AFAP1 at Ser277, PI3K activity enables its PH1 domain-dependent colocalization with c-Src, and its SH3-binding motif engages the c-Src SH3 domain to drive kinase activation, podosome formation, focal adhesion regulation, and cell migration; these activities are regulated by an intramolecular auto-inhibitory mechanism in which the carboxy-terminal leucine zipper constrains multimerization and actin cross-linking, and by Src-induced conformational changes that relieve this inhibition; additionally, AFAP1 mediates inflammatory (TNF-α/PKC) attenuation of P-glycoprotein efflux at the blood-brain barrier through a non-transcriptional phosphorylation-dependent mechanism."},"narrative":{"mechanistic_narrative":"AFAP1 (AFAP-110) is a multidomain adaptor and actin cross-linking protein that couples PKCα and PI3K signaling to c-Src activation and actin cytoskeletal remodeling, driving podosome formation, focal adhesion regulation, and cell migration [PMID:15314167, PMID:17360811]. Its amino terminus carries SH3- and SH2-binding modules that engage activated Src: an SH3-binding motif mediates stable complex formation and efficient tyrosine phosphorylation, while two independently functional SH2-binding regions (Y94 and Y451/Y453) cooperate in a multistep binding mechanism [PMID:9350057, PMID:9655255]. The carboxy-terminal actin-binding domain is necessary and sufficient for direct, cooperative actin filament binding and cross-linking, and is required for cytoskeletal localization [PMID:10666339, PMID:12134071, PMID:21984596]. These activities are held under intramolecular auto-inhibition by a carboxy-terminal leucine zipper that contacts the PH1 domain and constrains multimerization; structural disruption of the zipper, or activated-Src-induced conformational change, relieves this inhibition and separates the distinct outputs of actin cross-linking versus Src activation [PMID:9619827, PMID:14755689]. Upstream, PKCα phosphorylates AFAP1 at Ser277 to regulate podosome stability and turnover, PI3K activity and the PH1 domain enable AFAP1–c-Src colocalization, and Rho acts downstream in the actin reorganization arm [PMID:11641786, PMID:17360811, PMID:18577577]. AFAP1 is required for stress fiber and focal adhesion formation, integrin β1-dependent adhesion, and tumorigenic growth of prostate and breast cancer cells [PMID:17520695, PMID:17885682], and it is recruited to podosomes by the Tks5 fifth SH3 domain [PMID:19540230]. Independently, AFAP1 is a phosphorylation-dependent, non-transcriptional mediator of TNF-α–induced attenuation of P-glycoprotein efflux at the blood-brain barrier [PMID:28112407].","teleology":[{"year":1995,"claim":"Identifying a brain-specific alternatively spliced variant established that the AFAP locus encodes multiple proteins and revealed protein-interaction modules through a novel internal proline-rich SH3-binding sequence.","evidence":"cDNA cloning, northern/western blot, and GST-NINS pulldown of a 67-kDa partner in tissue lysates","pmids":["7876134"],"confidence":"Medium","gaps":["Identity of the 67-kDa interacting protein not determined","Functional role of the AFAP-120 variant not established"]},{"year":1997,"claim":"Mapping the amino-terminal SH3-binding motif answered how AFAP-110 docks onto Src, showing this motif is required for stable Src complex formation and efficient tyrosine phosphorylation.","evidence":"Site-directed mutagenesis, GST-SH3 pulldown, and co-IP with activated Src527F in Cos-1 cells","pmids":["9350057"],"confidence":"High","gaps":["Did not address the contribution of SH2-binding tyrosines to complex formation","Consequences for downstream actin remodeling not tested"]},{"year":1998,"claim":"Defining two SH2-binding tyrosine regions and a self-associating leucine zipper resolved the architecture of Src binding and revealed activated Src remodels AFAP-110 into multimers.","evidence":"Tyrosine mutagenesis with co-IP/phosphorylation analysis, plus GST affinity absorption, size-exclusion chromatography, and deletional mutagenesis with imaging in Src527F cells","pmids":["9655255","9619827"],"confidence":"High","gaps":["Mechanism coupling multimer state to Src activation not yet defined","In vitro reconstitution of the conformational switch not performed"]},{"year":2000,"claim":"Localizing direct actin-filament binding to the carboxy terminus, and showing leucine zipper deletion unleashes actin-altering activity, established that the leucine zipper auto-inhibits AFAP-110's intrinsic cytoskeletal function.","evidence":"In vitro actin binding with recombinant C-terminal domain, deletional mutagenesis, and immunofluorescence in untransformed fibroblasts","pmids":["10666339"],"confidence":"High","gaps":["Whether actin alteration requires kinase activation not resolved here","Structural basis of auto-inhibition not defined"]},{"year":2001,"claim":"Epistasis experiments placed AFAP-110-directed Src activation upstream of Rho-dependent actin reorganization and showed the SH3-binding motif and PH domain are required for these activities.","evidence":"Expression of Δlzip and point mutants, kinase assays, and co-expression with dominant-active RhoA(V14)","pmids":["11641786"],"confidence":"High","gaps":["Direct Rho activation mechanism by AFAP-110 not shown","Physiological trigger for the conformational change not identified"]},{"year":2002,"claim":"Reconstituting cooperative actin cross-linking and showing PKC phosphorylation alters AFAP-110 conformation linked an upstream kinase to its cytoskeletal cross-linking function.","evidence":"In vitro actin cross-linking and PKC phosphorylation of recombinant AFAP-110, size-exclusion chromatography, and PMA treatment with a PKC-binding-deficient mutant","pmids":["12134071"],"confidence":"High","gaps":["Specific PKC phosphorylation site not yet mapped","Quantitative relationship between multimer destabilization and cross-linking not defined"]},{"year":2004,"claim":"Two studies established that PKCα signaling requires AFAP-110 to activate c-Src and form podosomes, and dissected the leucine zipper into separable sequence (cross-linking) and structural (Src activation) determinants contacting the PH1 domain.","evidence":"Rescue with domain mutants in AFAP-110-deficient CaOV3 cells, plus leucine zipper substitution/point mutagenesis with in vitro cross-linking and cellular cSrc activation assays","pmids":["15314167","14755689"],"confidence":"High","gaps":["Atomic structure of the PH1–leucine zipper auto-inhibitory contact not solved","How PKCα binding triggers the conformational relief not detailed"]},{"year":2007,"claim":"Knockdown and in vivo tumor studies established AFAP1 as required for stress fiber and focal adhesion formation, integrin β1-dependent adhesion, migration, and tumorigenic growth, with PI3K positioned upstream of AFAP1–c-Src colocalization.","evidence":"siRNA/inducible knockdown with adhesion and imaging assays in breast cancer cells; domain-mutant rescue plus orthotopic mouse model in prostate cancer cells; PI3K inhibition/p85 knockout with wound-healing assay","pmids":["17520695","17885682","17360811"],"confidence":"High","gaps":["Direct link between PI3K lipid products and the PH1 domain not biochemically demonstrated","How AFAP1 loss reduces integrin β1 expression not mechanistically explained"]},{"year":2008,"claim":"Identifying Ser277 as the PKCα phosphorylation site and showing the S277A mutant increases podosome number and lifespan defined a regulatory phosphorylation controlling podosome turnover.","evidence":"Phospho-specific antibody, mutagenesis, and live-cell video microscopy of podosome dynamics in A7r5 cells","pmids":["18577577"],"confidence":"High","gaps":["Downstream effectors reading the Ser277 phosphorylation state not identified","Relationship of Ser277 to the leucine zipper/PH1 auto-inhibition not tested"]},{"year":2009,"claim":"Mislocalization experiments showed Tks5 recruits AFAP-110 (with p190RhoGAP and cortactin) to podosomes via its fifth SH3 domain, defining a scaffold that positions AFAP1 at invadosomal structures.","evidence":"Forced mitochondrial mislocalization of Tks5 deletion mutants, immunofluorescence, and siRNA knockdown","pmids":["19540230"],"confidence":"High","gaps":["Whether Tks5–AFAP1 binding is direct not established","Hierarchy of recruitment among AFAP1, cortactin and p190RhoGAP unresolved"]},{"year":2011,"claim":"Fractionation, ABD deletion, and paralog comparison clarified that actin-bound localization governs AFAP1 cytoskeletal function and Src engagement, and that AFAP1 selectively binds c-Src SH3 (versus cortactin) distinguishing it from AFAP1L1.","evidence":"Subcellular fractionation with actin-depolymerizing drugs and ABD-deletion imaging/gelatin assay; SH3-domain binding specificity comparison and overexpression in A7r5 cells","pmids":["21984596","21333378"],"confidence":"High","gaps":["Functional consequence of AFAP1 selectivity for c-Src over cortactin in vivo not tested","Whether ABD-dependent localization is regulated by signaling not addressed"]},{"year":2017,"claim":"Phosphoproteomics and selective knockdown identified AFAP1 as a non-transcriptional mediator of TNF-α–induced attenuation of P-glycoprotein efflux at the blood-brain barrier, extending AFAP1 function beyond cytoskeletal/Src signaling.","evidence":"Quantitative phosphoproteomics, siRNA knockdown with P-gp efflux assay, negative controls (MAPK1, AP-1), and in vivo LPS mouse model","pmids":["28112407"],"confidence":"High","gaps":["Phosphorylation site(s) on AFAP1 driving P-gp modulation not mapped","Mechanism by which phospho-AFAP1 reduces P-gp transport activity unknown"]},{"year":null,"claim":"How AFAP1's well-defined cytoskeletal/Src adaptor activities mechanistically connect to its non-transcriptional regulation of P-glycoprotein at the blood-brain barrier remains unresolved.","evidence":"","pmids":[],"confidence":"High","gaps":["No structure of full-length AFAP1 or its auto-inhibited state","The effector linking phospho-AFAP1 to P-gp transport activity is unidentified","Direct biochemical link between PI3K products and the PH1 domain not demonstrated"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0060090","term_label":"molecular adaptor activity","supporting_discovery_ids":[0,1,11]},{"term_id":"GO:0008092","term_label":"cytoskeletal protein binding","supporting_discovery_ids":[4,6,15]},{"term_id":"GO:0005198","term_label":"structural molecule activity","supporting_discovery_ids":[6,9]},{"term_id":"GO:0098772","term_label":"molecular function regulator activity","supporting_discovery_ids":[0,5,8]}],"localization":[{"term_id":"GO:0005856","term_label":"cytoskeleton","supporting_discovery_ids":[4,6,15]},{"term_id":"GO:0005829","term_label":"cytosol","supporting_discovery_ids":[15]},{"term_id":"GO:0005886","term_label":"plasma membrane","supporting_discovery_ids":[3,15]}],"pathway":[{"term_id":"R-HSA-162582","term_label":"Signal Transduction","supporting_discovery_ids":[0,5,11]},{"term_id":"R-HSA-1474244","term_label":"Extracellular matrix organization","supporting_discovery_ids":[9,10]},{"term_id":"R-HSA-1643685","term_label":"Disease","supporting_discovery_ids":[10,17]}],"complexes":["podosome","focal adhesion"],"partners":["SRC","PRKCA","TKS5","CTTN","PIK3R1"],"other_free_text":[]}},"prefetch_data":{"uniprot":{"accession":"Q8N556","full_name":"Actin filament-associated protein 1","aliases":["110 kDa actin filament-associated protein","AFAP-110"],"length_aa":730,"mass_kda":80.7,"function":"Can cross-link actin filaments into both network and bundle structures (By similarity). May modulate changes in actin filament integrity and induce lamellipodia formation. May function as an adapter molecule that links other proteins, such as SRC and PKC to the actin cytoskeleton. Seems to play a role in the development and progression of prostate adenocarcinoma by regulating cell-matrix adhesions and migration in the cancer cells","subcellular_location":"Cytoplasm, cytoskeleton, stress fiber","url":"https://www.uniprot.org/uniprotkb/Q8N556/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/AFAP1","classification":"Not Classified","n_dependent_lines":0,"n_total_lines":1208,"dependency_fraction":0.0},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[{"gene":"CSNK2B","stoichiometry":0.2}],"url":"https://opencell.sf.czbiohub.org/search/AFAP1","total_profiled":1310},"omim":[{"mim_id":"619779","title":"AFAP1 ANTISENSE RNA 1, NONCODING; AFAP1AS1","url":"https://www.omim.org/entry/619779"},{"mim_id":"614410","title":"ACTIN FILAMENT-ASSOCIATED PROTEIN 1-LIKE 1; AFAP1L1","url":"https://www.omim.org/entry/614410"},{"mim_id":"612420","title":"ACTIN FILAMENT-ASSOCIATED PROTEIN 1-LIKE PROTEIN 2; AFAP1L2","url":"https://www.omim.org/entry/612420"},{"mim_id":"608252","title":"ACTIN FILAMENT-ASSOCIATED PROTEIN 1; AFAP1","url":"https://www.omim.org/entry/608252"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"Supported","locations":[{"location":"Actin filaments","reliability":"Supported"},{"location":"Focal adhesion sites","reliability":"Additional"},{"location":"Cytosol","reliability":"Additional"}],"tissue_specificity":"Low tissue specificity","tissue_distribution":"Detected in many","driving_tissues":[],"url":"https://www.proteinatlas.org/search/AFAP1"},"hgnc":{"alias_symbol":["AFAP-110","AFAP"],"prev_symbol":[]},"alphafold":{"accession":"Q8N556","domains":[{"cath_id":"2.30.29.30","chopping":"155-250","consensus_level":"high","plddt":88.3452,"start":155,"end":250},{"cath_id":"2.30.29.30","chopping":"352-455","consensus_level":"high","plddt":86.7821,"start":352,"end":455},{"cath_id":"1.10.12","chopping":"2-42","consensus_level":"high","plddt":86.8154,"start":2,"end":42},{"cath_id":"1.10.287","chopping":"561-649","consensus_level":"high","plddt":91.973,"start":561,"end":649}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/Q8N556","model_url":"https://alphafold.ebi.ac.uk/files/AF-Q8N556-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-Q8N556-F1-predicted_aligned_error_v6.png","plddt_mean":65.81},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=AFAP1","jax_strain_url":"https://www.jax.org/strain/search?query=AFAP1"},"sequence":{"accession":"Q8N556","fasta_url":"https://rest.uniprot.org/uniprotkb/Q8N556.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/Q8N556/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/Q8N556"}},"corpus_meta":[{"pmid":"23333711","id":"PMC_23333711","title":"Hypomethylation of noncoding DNA regions and overexpression of the long noncoding RNA, AFAP1-AS1, in Barrett's esophagus and esophageal adenocarcinoma.","date":"2013","source":"Gastroenterology","url":"https://pubmed.ncbi.nlm.nih.gov/23333711","citation_count":206,"is_preprint":false},{"pmid":"14574166","id":"PMC_14574166","title":"Attenuated familial adenomatous polyposis (AFAP). A review of the literature.","date":"2003","source":"Familial cancer","url":"https://pubmed.ncbi.nlm.nih.gov/14574166","citation_count":198,"is_preprint":false},{"pmid":"8625067","id":"PMC_8625067","title":"Attenuated familial adenomatous polyposis (AFAP). A phenotypically and genotypically distinctive variant of FAP.","date":"1995","source":"Cancer","url":"https://pubmed.ncbi.nlm.nih.gov/8625067","citation_count":190,"is_preprint":false},{"pmid":"32020881","id":"PMC_32020881","title":"Exosome-mediated lncRNA AFAP1-AS1 promotes trastuzumab resistance through binding with AUF1 and activating ERBB2 translation.","date":"2020","source":"Molecular cancer","url":"https://pubmed.ncbi.nlm.nih.gov/32020881","citation_count":151,"is_preprint":false},{"pmid":"30326930","id":"PMC_30326930","title":"Long noncoding RNA AFAP1-AS1 acts as a competing endogenous RNA of miR-423-5p to facilitate nasopharyngeal carcinoma metastasis through regulating the Rho/Rac pathway.","date":"2018","source":"Journal of experimental & clinical cancer research : CR","url":"https://pubmed.ncbi.nlm.nih.gov/30326930","citation_count":144,"is_preprint":false},{"pmid":"26245991","id":"PMC_26245991","title":"AFAP1-AS1, a long noncoding RNA upregulated in lung cancer and promotes invasion and metastasis.","date":"2015","source":"Tumour biology : the journal of the International Society for Oncodevelopmental Biology and Medicine","url":"https://pubmed.ncbi.nlm.nih.gov/26245991","citation_count":135,"is_preprint":false},{"pmid":"33230475","id":"PMC_33230475","title":"M2 Macrophage-Derived Exosomal lncRNA AFAP1-AS1 and MicroRNA-26a Affect Cell Migration and Metastasis in Esophageal Cancer.","date":"2020","source":"Molecular therapy. Nucleic acids","url":"https://pubmed.ncbi.nlm.nih.gov/33230475","citation_count":112,"is_preprint":false},{"pmid":"32999837","id":"PMC_32999837","title":"Nanoparticles (NPs)-Meditated LncRNA AFAP1-AS1 Silencing to Block Wnt/β-Catenin Signaling Pathway for Synergistic Reversal of Radioresistance and Effective Cancer Radiotherapy.","date":"2020","source":"Advanced science (Weinheim, Baden-Wurttemberg, Germany)","url":"https://pubmed.ncbi.nlm.nih.gov/32999837","citation_count":86,"is_preprint":false},{"pmid":"31443665","id":"PMC_31443665","title":"LncRNA AFAP1-AS1 promotes tumorigenesis and epithelial-mesenchymal transition of osteosarcoma through RhoC/ROCK1/p38MAPK/Twist1 signaling pathway.","date":"2019","source":"Journal of experimental & clinical cancer research : CR","url":"https://pubmed.ncbi.nlm.nih.gov/31443665","citation_count":85,"is_preprint":false},{"pmid":"15314167","id":"PMC_15314167","title":"Protein kinase Calpha activates c-Src and induces podosome formation via AFAP-110.","date":"2004","source":"Molecular and cellular biology","url":"https://pubmed.ncbi.nlm.nih.gov/15314167","citation_count":85,"is_preprint":false},{"pmid":"34168109","id":"PMC_34168109","title":"Long non-coding RNA AFAP1-AS1 accelerates lung cancer cells migration and invasion by interacting with SNIP1 to upregulate c-Myc.","date":"2021","source":"Signal transduction and targeted therapy","url":"https://pubmed.ncbi.nlm.nih.gov/34168109","citation_count":78,"is_preprint":false},{"pmid":"31696057","id":"PMC_31696057","title":"LncRNA AFAP1-AS1 Supresses miR-139-5p and Promotes Cell Proliferation and Chemotherapy Resistance of Non-small Cell Lung Cancer by Competitively Upregulating RRM2.","date":"2019","source":"Frontiers in oncology","url":"https://pubmed.ncbi.nlm.nih.gov/31696057","citation_count":74,"is_preprint":false},{"pmid":"27577754","id":"PMC_27577754","title":"AFAP1-AS1 is upregulated and promotes esophageal squamous cell carcinoma cell proliferation and inhibits cell apoptosis.","date":"2016","source":"Cancer medicine","url":"https://pubmed.ncbi.nlm.nih.gov/27577754","citation_count":67,"is_preprint":false},{"pmid":"29057544","id":"PMC_29057544","title":"AFAP1-AS1: A novel oncogenic long non-coding RNA in human cancers.","date":"2017","source":"Cell proliferation","url":"https://pubmed.ncbi.nlm.nih.gov/29057544","citation_count":60,"is_preprint":false},{"pmid":"11607843","id":"PMC_11607843","title":"The actin filament-associated protein AFAP-110 is an adaptor protein that modulates changes in actin filament integrity.","date":"2001","source":"Oncogene","url":"https://pubmed.ncbi.nlm.nih.gov/11607843","citation_count":58,"is_preprint":false},{"pmid":"29310682","id":"PMC_29310682","title":"Upregulation of the long non-coding RNA AFAP1-AS1 affects the proliferation, invasion and survival of tongue squamous cell carcinoma via the Wnt/β-catenin signaling pathway.","date":"2018","source":"Molecular cancer","url":"https://pubmed.ncbi.nlm.nih.gov/29310682","citation_count":58,"is_preprint":false},{"pmid":"28938565","id":"PMC_28938565","title":"LncRNA AFAP1-AS1 promotes growth and metastasis of cholangiocarcinoma cells.","date":"2017","source":"Oncotarget","url":"https://pubmed.ncbi.nlm.nih.gov/28938565","citation_count":57,"is_preprint":false},{"pmid":"28535506","id":"PMC_28535506","title":"Long Noncoding RNA AFAP1-AS1 Promoted Tumor Growth and Invasion in Cholangiocarcinoma.","date":"2017","source":"Cellular physiology and biochemistry : international journal of experimental cellular physiology, biochemistry, and pharmacology","url":"https://pubmed.ncbi.nlm.nih.gov/28535506","citation_count":56,"is_preprint":false},{"pmid":"17885682","id":"PMC_17885682","title":"AFAP-110 is overexpressed in prostate cancer and contributes to tumorigenic growth by regulating focal contacts.","date":"2007","source":"The Journal of clinical investigation","url":"https://pubmed.ncbi.nlm.nih.gov/17885682","citation_count":56,"is_preprint":false},{"pmid":"28451917","id":"PMC_28451917","title":"Long Noncoding RNA AFAP1-AS1 Promotes Cell Proliferation and Apoptosis of Gastric Cancer Cells via PTEN/p-AKT Pathway.","date":"2017","source":"Digestive diseases and sciences","url":"https://pubmed.ncbi.nlm.nih.gov/28451917","citation_count":54,"is_preprint":false},{"pmid":"30505272","id":"PMC_30505272","title":"AFAP1-AS1 Promotes Epithelial-Mesenchymal Transition and Tumorigenesis Through Wnt/β-Catenin Signaling Pathway in Triple-Negative Breast Cancer.","date":"2018","source":"Frontiers in pharmacology","url":"https://pubmed.ncbi.nlm.nih.gov/30505272","citation_count":53,"is_preprint":false},{"pmid":"26803513","id":"PMC_26803513","title":"Critical role for the long non-coding RNA AFAP1-AS1 in the proliferation and metastasis of hepatocellular carcinoma.","date":"2016","source":"Tumour biology : the journal of the International Society for Oncodevelopmental Biology and Medicine","url":"https://pubmed.ncbi.nlm.nih.gov/26803513","citation_count":53,"is_preprint":false},{"pmid":"36871154","id":"PMC_36871154","title":"The Tumorigenic Effect of lncRNA AFAP1-AS1 is Mediated by Translated Peptide ATMLP Under the Control of m6 A Methylation.","date":"2023","source":"Advanced science (Weinheim, Baden-Wurttemberg, Germany)","url":"https://pubmed.ncbi.nlm.nih.gov/36871154","citation_count":52,"is_preprint":false},{"pmid":"30173945","id":"PMC_30173945","title":"The role of long non-coding RNA AFAP1-AS1 in human malignant tumors.","date":"2018","source":"Pathology, research and practice","url":"https://pubmed.ncbi.nlm.nih.gov/30173945","citation_count":52,"is_preprint":false},{"pmid":"19540230","id":"PMC_19540230","title":"Tks5 recruits AFAP-110, p190RhoGAP, and cortactin for podosome formation.","date":"2009","source":"Experimental cell research","url":"https://pubmed.ncbi.nlm.nih.gov/19540230","citation_count":51,"is_preprint":false},{"pmid":"19793053","id":"PMC_19793053","title":"APC or MUTYH mutations account for the majority of clinically well-characterized families with FAP and AFAP phenotype and patients with more than 30 adenomas.","date":"2009","source":"Clinical genetics","url":"https://pubmed.ncbi.nlm.nih.gov/19793053","citation_count":50,"is_preprint":false},{"pmid":"27578191","id":"PMC_27578191","title":"Long non-coding RNA AFAP1-AS1 facilitates tumor growth and promotes metastasis in colorectal cancer.","date":"2016","source":"Biological research","url":"https://pubmed.ncbi.nlm.nih.gov/27578191","citation_count":49,"is_preprint":false},{"pmid":"9350057","id":"PMC_9350057","title":"The integrity of the SH3 binding motif of AFAP-110 is required to facilitate tyrosine phosphorylation by, and stable complex formation with, Src.","date":"1997","source":"Molecular and cellular biochemistry","url":"https://pubmed.ncbi.nlm.nih.gov/9350057","citation_count":47,"is_preprint":false},{"pmid":"9619827","id":"PMC_9619827","title":"Src can regulate carboxy terminal interactions with AFAP-110, which influence self-association, cell localization and actin filament integrity.","date":"1998","source":"Oncogene","url":"https://pubmed.ncbi.nlm.nih.gov/9619827","citation_count":46,"is_preprint":false},{"pmid":"29971915","id":"PMC_29971915","title":"Long non-coding RNA AFAP1-AS1/miR-320a/RBPJ axis regulates laryngeal carcinoma cell stemness and chemoresistance.","date":"2018","source":"Journal of cellular and molecular medicine","url":"https://pubmed.ncbi.nlm.nih.gov/29971915","citation_count":45,"is_preprint":false},{"pmid":"17520695","id":"PMC_17520695","title":"AFAP-110 is required for actin stress fiber formation and cell adhesion in MDA-MB-231 breast cancer cells.","date":"2007","source":"Journal of cellular physiology","url":"https://pubmed.ncbi.nlm.nih.gov/17520695","citation_count":44,"is_preprint":false},{"pmid":"10666339","id":"PMC_10666339","title":"The carboxy terminus of AFAP-110 modulates direct interactions with actin filaments and regulates its ability to alter actin filament integrity and induce lamellipodia formation.","date":"2000","source":"Experimental cell research","url":"https://pubmed.ncbi.nlm.nih.gov/10666339","citation_count":44,"is_preprint":false},{"pmid":"30278439","id":"PMC_30278439","title":"lncRNA AFAP1-AS1 Promotes Migration and Invasion of Non-Small Cell Lung Cancer via Up-Regulating IRF7 and the RIG-I-Like Receptor Signaling Pathway.","date":"2018","source":"Cellular physiology and biochemistry : international journal of experimental cellular physiology, biochemistry, and pharmacology","url":"https://pubmed.ncbi.nlm.nih.gov/30278439","citation_count":44,"is_preprint":false},{"pmid":"32376943","id":"PMC_32376943","title":"lncRNA AFAP1-AS1 promotes triple negative breast cancer cell proliferation and invasion via targeting miR-145 to regulate MTH1 expression.","date":"2020","source":"Scientific reports","url":"https://pubmed.ncbi.nlm.nih.gov/32376943","citation_count":43,"is_preprint":false},{"pmid":"30293090","id":"PMC_30293090","title":"Long non-coding RNA AFAP1-AS1 plays an oncogenic role in promoting cell migration in non-small cell lung cancer.","date":"2018","source":"Cellular and molecular life sciences : CMLS","url":"https://pubmed.ncbi.nlm.nih.gov/30293090","citation_count":42,"is_preprint":false},{"pmid":"11641786","id":"PMC_11641786","title":"The intrinsic ability of AFAP-110 to alter actin filament integrity is linked with its ability to also activate cellular tyrosine kinases.","date":"2001","source":"Oncogene","url":"https://pubmed.ncbi.nlm.nih.gov/11641786","citation_count":41,"is_preprint":false},{"pmid":"32228518","id":"PMC_32228518","title":"Long non-coding RNA AFAP1-AS1 accelerates the progression of melanoma by targeting miR-653-5p/RAI14 axis.","date":"2020","source":"BMC cancer","url":"https://pubmed.ncbi.nlm.nih.gov/32228518","citation_count":40,"is_preprint":false},{"pmid":"23561487","id":"PMC_23561487","title":"Mutational spectrum of the APC and MUTYH genes and genotype-phenotype correlations in Brazilian FAP, AFAP, and MAP patients.","date":"2013","source":"Orphanet journal of rare diseases","url":"https://pubmed.ncbi.nlm.nih.gov/23561487","citation_count":40,"is_preprint":false},{"pmid":"12134071","id":"PMC_12134071","title":"PC phosphorylation increases the ability of AFAP-110 to cross-link actin filaments.","date":"2002","source":"Molecular biology of the cell","url":"https://pubmed.ncbi.nlm.nih.gov/12134071","citation_count":39,"is_preprint":false},{"pmid":"17360811","id":"PMC_17360811","title":"PI3K activation is required for PMA-directed activation of cSrc by AFAP-110.","date":"2007","source":"American journal of physiology. Cell physiology","url":"https://pubmed.ncbi.nlm.nih.gov/17360811","citation_count":38,"is_preprint":false},{"pmid":"34145213","id":"PMC_34145213","title":"Long noncoding RNA AFAP1-AS1 promotes tumor progression and invasion by regulating the miR-2110/Sp1 axis in triple-negative breast cancer.","date":"2021","source":"Cell death & disease","url":"https://pubmed.ncbi.nlm.nih.gov/34145213","citation_count":37,"is_preprint":false},{"pmid":"21333378","id":"PMC_21333378","title":"AFAP1L1 is a novel adaptor protein of the AFAP family that interacts with cortactin and localizes to invadosomes.","date":"2011","source":"European journal of cell biology","url":"https://pubmed.ncbi.nlm.nih.gov/21333378","citation_count":37,"is_preprint":false},{"pmid":"28670387","id":"PMC_28670387","title":"Down-regulation of long non-coding RNA AFAP1-AS1 inhibits tumor cell growth and invasion in lung adenocarcinoma.","date":"2017","source":"American journal of translational research","url":"https://pubmed.ncbi.nlm.nih.gov/28670387","citation_count":37,"is_preprint":false},{"pmid":"30300116","id":"PMC_30300116","title":"The long coding RNA AFAP1-AS1 promotes tumor cell growth and invasion in pancreatic cancer through upregulating the IGF1R oncogene via sequestration of miR-133a.","date":"2018","source":"Cell cycle (Georgetown, Tex.)","url":"https://pubmed.ncbi.nlm.nih.gov/30300116","citation_count":34,"is_preprint":false},{"pmid":"30506548","id":"PMC_30506548","title":"Long non-coding RNA AFAP1-AS1 promoting epithelial-mesenchymal transition of endometriosis is correlated with transcription factor ZEB1.","date":"2018","source":"American journal of reproductive immunology (New York, N.Y. : 1989)","url":"https://pubmed.ncbi.nlm.nih.gov/30506548","citation_count":33,"is_preprint":false},{"pmid":"29080690","id":"PMC_29080690","title":"Circulating long non-coding RNA AFAP1-AS1 is a potential diagnostic biomarker for non-small cell lung cancer.","date":"2017","source":"Clinica chimica acta; international journal of clinical chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/29080690","citation_count":33,"is_preprint":false},{"pmid":"30819221","id":"PMC_30819221","title":"Cross-talk among AFAP1-AS1, ACVR1 and microRNA-384 regulates the stemness of pancreatic cancer cells and tumorigenicity in nude mice.","date":"2019","source":"Journal of experimental & clinical cancer research : CR","url":"https://pubmed.ncbi.nlm.nih.gov/30819221","citation_count":32,"is_preprint":false},{"pmid":"30854123","id":"PMC_30854123","title":"Cloning and characterization of the putative AFAP1-AS1 promoter region.","date":"2019","source":"Journal of Cancer","url":"https://pubmed.ncbi.nlm.nih.gov/30854123","citation_count":30,"is_preprint":false},{"pmid":"30206930","id":"PMC_30206930","title":"Cucurbitacin B suppresses proliferation of pancreatic cancer cells by ceRNA: Effect of miR-146b-5p and lncRNA-AFAP1-AS1.","date":"2018","source":"Journal of cellular physiology","url":"https://pubmed.ncbi.nlm.nih.gov/30206930","citation_count":30,"is_preprint":false},{"pmid":"29254250","id":"PMC_29254250","title":"Long non-coding RNA AFAP1-AS1 is a novel biomarker in various cancers: a systematic review and meta-analysis based on the literature and GEO datasets.","date":"2017","source":"Oncotarget","url":"https://pubmed.ncbi.nlm.nih.gov/29254250","citation_count":30,"is_preprint":false},{"pmid":"9655255","id":"PMC_9655255","title":"Formation of a stable src-AFAP-110 complex through either an amino-terminal or a carboxy-terminal SH2-binding motif.","date":"1998","source":"Molecular carcinogenesis","url":"https://pubmed.ncbi.nlm.nih.gov/9655255","citation_count":29,"is_preprint":false},{"pmid":"33138677","id":"PMC_33138677","title":"LncRNA AFAP1-AS1 modulates the sensitivity of paclitaxel-resistant prostate cancer cells to paclitaxel via miR-195-5p/FKBP1A axis.","date":"2020","source":"Cancer biology & therapy","url":"https://pubmed.ncbi.nlm.nih.gov/33138677","citation_count":29,"is_preprint":false},{"pmid":"27471399","id":"PMC_27471399","title":"Long noncoding RNA AFAP1-AS1, a potential novel biomarker to predict the clinical outcome of cancer patients: a meta-analysis.","date":"2016","source":"OncoTargets and therapy","url":"https://pubmed.ncbi.nlm.nih.gov/27471399","citation_count":29,"is_preprint":false},{"pmid":"29888110","id":"PMC_29888110","title":"Long noncoding RNA AFAP1-AS1 facilitates tumor growth through enhancer of zeste homolog 2 in colorectal cancer.","date":"2018","source":"American journal of cancer research","url":"https://pubmed.ncbi.nlm.nih.gov/29888110","citation_count":29,"is_preprint":false},{"pmid":"18577577","id":"PMC_18577577","title":"Phosphorylation of AFAP-110 affects podosome lifespan in A7r5 cells.","date":"2008","source":"Journal of cell science","url":"https://pubmed.ncbi.nlm.nih.gov/18577577","citation_count":27,"is_preprint":false},{"pmid":"29331858","id":"PMC_29331858","title":"Down-regulation of long non-coding RNA AFAP1-AS1 inhibits tumor growth, promotes apoptosis and decreases metastasis in thyroid cancer.","date":"2018","source":"Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie","url":"https://pubmed.ncbi.nlm.nih.gov/29331858","citation_count":27,"is_preprint":false},{"pmid":"31897216","id":"PMC_31897216","title":"AFAP1-AS1 induces cisplatin resistance in non-small cell lung cancer through PI3K/AKT pathway.","date":"2019","source":"Oncology letters","url":"https://pubmed.ncbi.nlm.nih.gov/31897216","citation_count":26,"is_preprint":false},{"pmid":"32504788","id":"PMC_32504788","title":"LncRNA AFAP1-AS1 contributes to the progression of endometrial carcinoma by regulating miR-545-3p/VEGFA pathway.","date":"2020","source":"Molecular and cellular probes","url":"https://pubmed.ncbi.nlm.nih.gov/32504788","citation_count":26,"is_preprint":false},{"pmid":"31314060","id":"PMC_31314060","title":"Downregulation of lncRNA AFAP1-AS1 by oridonin inhibits the epithelial-to-mesenchymal transition and proliferation of pancreatic cancer cells.","date":"2019","source":"Acta biochimica et biophysica Sinica","url":"https://pubmed.ncbi.nlm.nih.gov/31314060","citation_count":26,"is_preprint":false},{"pmid":"32280542","id":"PMC_32280542","title":"Bidirectional interaction of lncRNA AFAP1-AS1 and CRKL accelerates the proliferative and metastatic abilities of hepatocarcinoma cells.","date":"2020","source":"Journal of advanced research","url":"https://pubmed.ncbi.nlm.nih.gov/32280542","citation_count":25,"is_preprint":false},{"pmid":"28924375","id":"PMC_28924375","title":"LncRNA AFAP1-AS Functions as a Competing Endogenous RNA to Regulate RAP1B Expression by sponging miR-181a in the HSCR.","date":"2017","source":"International journal of medical sciences","url":"https://pubmed.ncbi.nlm.nih.gov/28924375","citation_count":25,"is_preprint":false},{"pmid":"34289449","id":"PMC_34289449","title":"Formononetin relieves the facilitating effect of lncRNA AFAP1-AS1-miR-195/miR-545 axis on progression and chemo-resistance of triple-negative breast cancer.","date":"2021","source":"Aging","url":"https://pubmed.ncbi.nlm.nih.gov/34289449","citation_count":25,"is_preprint":false},{"pmid":"33999777","id":"PMC_33999777","title":"LncRNA AFAP-AS1 promotes anaplastic thyroid cancer progression by sponging miR-155-5p through ETS1/ERK pathway.","date":"2021","source":"Bioengineered","url":"https://pubmed.ncbi.nlm.nih.gov/33999777","citation_count":24,"is_preprint":false},{"pmid":"31132513","id":"PMC_31132513","title":"LncRNA GAS8-AS inhibits colorectal cancer (CRC) cell proliferation by downregulating lncRNA AFAP1-AS1.","date":"2019","source":"Gene","url":"https://pubmed.ncbi.nlm.nih.gov/31132513","citation_count":23,"is_preprint":false},{"pmid":"30905507","id":"PMC_30905507","title":"The bZIP transcription factor Afap1 mediates the oxidative stress response and aflatoxin biosynthesis in Aspergillus flavus.","date":"2019","source":"Revista Argentina de microbiologia","url":"https://pubmed.ncbi.nlm.nih.gov/30905507","citation_count":23,"is_preprint":false},{"pmid":"33999309","id":"PMC_33999309","title":"AFAP1-AS1: a rising star among oncogenic long non-coding RNAs.","date":"2021","source":"Science China. Life sciences","url":"https://pubmed.ncbi.nlm.nih.gov/33999309","citation_count":22,"is_preprint":false},{"pmid":"28112407","id":"PMC_28112407","title":"Actin filament-associated protein 1 (AFAP-1) is a key mediator in inflammatory signaling-induced rapid attenuation of intrinsic P-gp function in human brain capillary endothelial cells.","date":"2017","source":"Journal of neurochemistry","url":"https://pubmed.ncbi.nlm.nih.gov/28112407","citation_count":22,"is_preprint":false},{"pmid":"29901121","id":"PMC_29901121","title":"Long noncoding RNA AFAP1‑AS1 enhances cell proliferation and invasion in osteosarcoma through regulating miR‑4695‑5p/TCF4‑β‑catenin signaling.","date":"2018","source":"Molecular medicine reports","url":"https://pubmed.ncbi.nlm.nih.gov/29901121","citation_count":21,"is_preprint":false},{"pmid":"31299308","id":"PMC_31299308","title":"AFAP1-AS1 Promotes Proliferation of Pituitary Adenoma Cells through miR-103a-3p to Activate PI3K/AKT Signaling Pathway.","date":"2019","source":"World neurosurgery","url":"https://pubmed.ncbi.nlm.nih.gov/31299308","citation_count":21,"is_preprint":false},{"pmid":"33949053","id":"PMC_33949053","title":"LncRNA AFAP1-AS1 regulates proliferation and apoptosis of endometriosis through activating STAT3/TGF-β/Smad signaling via miR-424-5p.","date":"2021","source":"The journal of obstetrics and gynaecology research","url":"https://pubmed.ncbi.nlm.nih.gov/33949053","citation_count":20,"is_preprint":false},{"pmid":"34335760","id":"PMC_34335760","title":"LncRNA AFAP1-AS1 Promotes the Progression of Colorectal Cancer through miR-195-5p and WISP1.","date":"2021","source":"Journal of oncology","url":"https://pubmed.ncbi.nlm.nih.gov/34335760","citation_count":20,"is_preprint":false},{"pmid":"35513844","id":"PMC_35513844","title":"Exosomal AFAP1-AS1 binds to microRNA-15a-5p to promote the proliferation, migration, and invasion of ectopic endometrial stromal cells in endometriosis.","date":"2022","source":"Reproductive biology and endocrinology : RB&E","url":"https://pubmed.ncbi.nlm.nih.gov/35513844","citation_count":20,"is_preprint":false},{"pmid":"14755689","id":"PMC_14755689","title":"Analysis of the role of the leucine zipper motif in regulating the ability of AFAP-110 to alter actin filament integrity.","date":"2004","source":"Journal of cellular biochemistry","url":"https://pubmed.ncbi.nlm.nih.gov/14755689","citation_count":20,"is_preprint":false},{"pmid":"32016968","id":"PMC_32016968","title":"Long non-coding RNA AFAP1-AS1 promotes proliferation and migration of gastric cancer by downregulating KLF2.","date":"2020","source":"European review for medical and pharmacological sciences","url":"https://pubmed.ncbi.nlm.nih.gov/32016968","citation_count":18,"is_preprint":false},{"pmid":"29439313","id":"PMC_29439313","title":"Expression analysis of AFAP1-AS1 and AFAP1 in breast cancer.","date":"2018","source":"Cancer biomarkers : section A of Disease markers","url":"https://pubmed.ncbi.nlm.nih.gov/29439313","citation_count":18,"is_preprint":false},{"pmid":"34043161","id":"PMC_34043161","title":"LncRNA AFAP1-AS1 promotes M1 polarization of macrophages and osteogenic differentiation of valve interstitial cells.","date":"2021","source":"Journal of physiology and biochemistry","url":"https://pubmed.ncbi.nlm.nih.gov/34043161","citation_count":18,"is_preprint":false},{"pmid":"35173146","id":"PMC_35173146","title":"Circ-AFAP1 promote clear cell renal cell carcinoma growth and angiogenesis by the Circ-AFAP1/miR-374b-3p/VEGFA signaling axis.","date":"2022","source":"Cell death discovery","url":"https://pubmed.ncbi.nlm.nih.gov/35173146","citation_count":18,"is_preprint":false},{"pmid":"28051261","id":"PMC_28051261","title":"Expression and functional role of long non-coding RNA AFAP1-AS1 in ovarian cancer.","date":"2016","source":"European review for medical and pharmacological sciences","url":"https://pubmed.ncbi.nlm.nih.gov/28051261","citation_count":18,"is_preprint":false},{"pmid":"33290312","id":"PMC_33290312","title":"The knockdown of LncRNA AFAP1-AS1 suppressed cell proliferation, migration, and invasion, and promoted apoptosis by regulating miR-545-3p/hepatoma-derived growth factor axis in lung cancer.","date":"2021","source":"Anti-cancer drugs","url":"https://pubmed.ncbi.nlm.nih.gov/33290312","citation_count":18,"is_preprint":false},{"pmid":"27862086","id":"PMC_27862086","title":"Correlations of AFAP1, GMDS and PTGFR gene polymorphisms with intra-ocular pressure response to latanoprost in patients with primary open-angle glaucoma.","date":"2016","source":"Journal of clinical pharmacy and therapeutics","url":"https://pubmed.ncbi.nlm.nih.gov/27862086","citation_count":18,"is_preprint":false},{"pmid":"38934786","id":"PMC_38934786","title":"FAP positive cancer-associated fibroblasts promote tumor progression and radioresistance in esophageal squamous cell carcinoma by transferring exosomal lncRNA AFAP1-AS1.","date":"2024","source":"Molecular carcinogenesis","url":"https://pubmed.ncbi.nlm.nih.gov/38934786","citation_count":17,"is_preprint":false},{"pmid":"29805596","id":"PMC_29805596","title":"Long non-coding RNA AFAP1-antisense RNA 1 promotes the proliferation, migration and invasion of gastric cancer cells and is associated with poor patient survival.","date":"2018","source":"Oncology letters","url":"https://pubmed.ncbi.nlm.nih.gov/29805596","citation_count":17,"is_preprint":false},{"pmid":"31669642","id":"PMC_31669642","title":"Long non-coding RNA AFAP1-AS1 promotes proliferation and invasion in prostate cancer via targeting miR-512-3p.","date":"2019","source":"Gene","url":"https://pubmed.ncbi.nlm.nih.gov/31669642","citation_count":15,"is_preprint":false},{"pmid":"31452798","id":"PMC_31452798","title":"Overexpression of lncRNA AFAP1-AS1 promotes cell proliferation and invasion in gastric cancer.","date":"2019","source":"Oncology letters","url":"https://pubmed.ncbi.nlm.nih.gov/31452798","citation_count":15,"is_preprint":false},{"pmid":"31785996","id":"PMC_31785996","title":"Circulatinglong non-coding RNA FEZF1-AS1 and AFAP1-AS1 serve as potential diagnostic biomarkers for gastric cancer.","date":"2019","source":"Pathology, research and practice","url":"https://pubmed.ncbi.nlm.nih.gov/31785996","citation_count":15,"is_preprint":false},{"pmid":"34334630","id":"PMC_34334630","title":"LncRNA AFAP1-AS1/miR-27b-3p/VEGF-C axis modulates stemness characteristics in cervical cancer cells.","date":"2021","source":"Chinese medical journal","url":"https://pubmed.ncbi.nlm.nih.gov/34334630","citation_count":15,"is_preprint":false},{"pmid":"31081081","id":"PMC_31081081","title":"Long noncoding RNA AFAP1-AS1 accelerates the proliferation and metastasis of prostate cancer via inhibiting RBM5 expression.","date":"2019","source":"European review for medical and pharmacological sciences","url":"https://pubmed.ncbi.nlm.nih.gov/31081081","citation_count":15,"is_preprint":false},{"pmid":"7876134","id":"PMC_7876134","title":"AFAP-120. A variant form of the Src SH2/SH3-binding partner AFAP-110 is detected in brain and contains a novel internal sequence which binds to a 67-kDa protein.","date":"1995","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/7876134","citation_count":14,"is_preprint":false},{"pmid":"29212176","id":"PMC_29212176","title":"Silencing of lncRNA AFAP1-AS1 suppressed lung cancer development by regulatory mechanism in cis and trans.","date":"2017","source":"Oncotarget","url":"https://pubmed.ncbi.nlm.nih.gov/29212176","citation_count":14,"is_preprint":false},{"pmid":"32801880","id":"PMC_32801880","title":"Silencing lncRNA AFAP1-AS1 Inhibits the Progression of Esophageal Squamous Cell Carcinoma Cells via Regulating the miR-498/VEGFA Axis.","date":"2020","source":"Cancer management and research","url":"https://pubmed.ncbi.nlm.nih.gov/32801880","citation_count":14,"is_preprint":false},{"pmid":"34126893","id":"PMC_34126893","title":"Long Non-coding RNA AFAP1-AS1 Facilitates Prostate Cancer Progression by Regulating miR-15b/IGF1R Axis.","date":"2021","source":"Current pharmaceutical design","url":"https://pubmed.ncbi.nlm.nih.gov/34126893","citation_count":13,"is_preprint":false},{"pmid":"32955920","id":"PMC_32955920","title":"LncRNA AFAP1-AS1 Knockdown Represses Cell Proliferation, Migration, and Induced Apoptosis in Breast Cancer by Downregulating SEPT2 Via Sponging miR-497-5p.","date":"2020","source":"Cancer biotherapy & radiopharmaceuticals","url":"https://pubmed.ncbi.nlm.nih.gov/32955920","citation_count":13,"is_preprint":false},{"pmid":"33330099","id":"PMC_33330099","title":"Long Noncoding RNA AFAP1-AS1 Is a Critical Regulator of Nasopharyngeal Carcinoma Tumorigenicity.","date":"2020","source":"Frontiers in oncology","url":"https://pubmed.ncbi.nlm.nih.gov/33330099","citation_count":13,"is_preprint":false},{"pmid":"30832752","id":"PMC_30832752","title":"Silencing of lncRNA AFAP1-AS1 Inhibits Cell Growth and Metastasis in Clear Cell Renal Cell Carcinoma.","date":"2019","source":"Oncology research","url":"https://pubmed.ncbi.nlm.nih.gov/30832752","citation_count":13,"is_preprint":false},{"pmid":"31938218","id":"PMC_31938218","title":"Knockdown of long non-coding RNA AFAP1-AS1 inhibits growth and promotes apoptosis in pituitary adenomas.","date":"2018","source":"International journal of clinical and experimental pathology","url":"https://pubmed.ncbi.nlm.nih.gov/31938218","citation_count":13,"is_preprint":false},{"pmid":"31945413","id":"PMC_31945413","title":"LncRNA AFAP1-AS1 promotes osteoblast differentiation of human aortic valve interstitial cells through regulating miR-155/SMAD5 axis.","date":"2020","source":"Molecular and cellular probes","url":"https://pubmed.ncbi.nlm.nih.gov/31945413","citation_count":13,"is_preprint":false},{"pmid":"32509208","id":"PMC_32509208","title":"Long noncoding RNA AFAP1-AS1 promotes osteosarcoma progression by regulating miR-497/IGF1R axis.","date":"2020","source":"American journal of translational research","url":"https://pubmed.ncbi.nlm.nih.gov/32509208","citation_count":13,"is_preprint":false},{"pmid":"21984596","id":"PMC_21984596","title":"The actin-binding domain of actin filament-associated protein (AFAP) is involved in the regulation of cytoskeletal structure.","date":"2011","source":"Cellular and molecular life sciences : CMLS","url":"https://pubmed.ncbi.nlm.nih.gov/21984596","citation_count":13,"is_preprint":false},{"pmid":"33926489","id":"PMC_33926489","title":"Long non-coding RNA AFAP1-AS1 facilitates ovarian cancer progression by regulating the miR-107/PDK4 axis.","date":"2021","source":"Journal of ovarian research","url":"https://pubmed.ncbi.nlm.nih.gov/33926489","citation_count":12,"is_preprint":false},{"pmid":"33650645","id":"PMC_33650645","title":"Long non‑coding RNA AFAP1‑AS1 facilitates the growth and invasiveness of oral squamous cell carcinoma by regulating the miR‑145/HOXA1 axis.","date":"2020","source":"Oncology reports","url":"https://pubmed.ncbi.nlm.nih.gov/33650645","citation_count":12,"is_preprint":false}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":57717,"output_tokens":5436,"usd":0.127346,"stage1_stop_reason":"end_turn"},"stage2":{"model":"claude-opus-4-8","input_tokens":13876,"output_tokens":4716,"usd":0.09364,"stage2_stop_reason":"end_turn"},"total_usd":0.220986,"stage1_batch_id":"msgbatch_0179Z2pxRYBsXi2xADneafRw","stage2_batch_id":"msgbatch_01LaY9ZaVdgjWArcZo9eqQE5","note":"batch pricing = 50% of standard"},"stage1_raw":"```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 2004,\n      \"finding\": \"AFAP-110 (AFAP1) is required to mediate PKCα activation of c-Src and subsequent podosome formation. PKCα activation directs AFAP-110 to colocalize with and bind the c-Src SH3 domain, activating the tyrosine kinase. In cells lacking AFAP-110 (CaOV3), PMA could not activate c-Src or induce podosomes; ectopic AFAP-110 rescued these effects. AFAP-110 mutants unable to bind or colocalize with c-Src failed to rescue.\",\n      \"method\": \"Immunofluorescence, ectopic expression of constitutively active PKCα, rescue experiments with AFAP-110 mutants in AFAP-110-deficient CaOV3 cells\",\n      \"journal\": \"Molecular and cellular biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — clean loss-of-function cell line + rescue with multiple domain mutants + defined cellular phenotype, single rigorous study\",\n      \"pmids\": [\"15314167\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2001,\n      \"finding\": \"AFAP-110 functions as an adaptor protein linking Src family kinases to actin filaments via: (1) SH3/SH2 binding modules in its amino terminus, (2) two pleckstrin homology domains, (3) a leucine zipper motif mediating self-association, (4) a carboxy-terminal actin-binding domain enabling direct actin filament interaction. Conformational changes associated with phosphorylation or mutagenesis reveal an intrinsic capability to alter actin filament integrity and activate cSrc.\",\n      \"method\": \"Review of prior biochemical, mutagenesis, and cell biology experiments including co-IP, pulldown, in vitro actin binding, and mutational analysis\",\n      \"journal\": \"Oncogene\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — synthesis of multiple orthogonal experimental approaches (pulldown, mutagenesis, in vitro binding, immunofluorescence) replicated across multiple papers\",\n      \"pmids\": [\"11607843\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 1997,\n      \"finding\": \"The amino-terminal SH3 binding motif I (SH3bm I) of AFAP-110 is required for stable complex formation with Src and for efficient tyrosine phosphorylation by Src. A proline-to-alanine mutation in SH3bm I abolished GST-SH3 absorption and reduced tyrosine phosphorylation ~2.7-fold, and prevented stable complex formation with activated Src (pp60-527F).\",\n      \"method\": \"Site-directed mutagenesis of SH3 binding motif, GST-SH3 pulldown, co-expression of Src527F with AFAP-110 mutants in Cos-1 cells, co-immunoprecipitation\",\n      \"journal\": \"Molecular and cellular biochemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — in vitro pulldown + mutagenesis + co-IP, single lab with multiple orthogonal methods\",\n      \"pmids\": [\"9350057\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 1998,\n      \"finding\": \"The carboxy-terminal 127 amino acids of AFAP-110 contain an alpha-helical leucine zipper motif required for self-association (multimerization). Activated Src (Src527F) disrupts carboxy-terminal interactions, shifting AFAP-110 to a multimeric population. Deletion of the leucine zipper region repositions actin filaments into rosette-like structures similar to Src-transformed cells and promotes lamellipodia-like structures. The carboxy terminus is required for colocalization with actin filaments and the cell membrane.\",\n      \"method\": \"GST fusion protein affinity absorption, Superose size-exclusion chromatography, deletional mutagenesis, immunofluorescence in cells co-expressing Src527F\",\n      \"journal\": \"Oncogene\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — in vitro pulldown reconstitution + size-exclusion chromatography + mutagenesis + cellular imaging, single lab multiple methods\",\n      \"pmids\": [\"9619827\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2000,\n      \"finding\": \"The carboxy terminus of AFAP-110 is both necessary and sufficient for direct actin filament association in vivo and in vitro. The carboxy-terminal actin-binding domain shows ~40% similarity to known actin-binding motifs. Deletion of the leucine zipper motif (AFAP-Δlzip) enables AFAP-110 to alter actin filament integrity and induce lamellipodia formation in untransformed cells. The leucine zipper motif normally constrains these activities.\",\n      \"method\": \"In vitro actin filament binding assay with recombinant carboxy-terminal domain, deletional mutagenesis, cellular expression with immunofluorescence in untransformed fibroblasts\",\n      \"journal\": \"Experimental cell research\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — in vitro actin binding reconstitution + mutagenesis + cellular phenotype, single lab with multiple orthogonal methods\",\n      \"pmids\": [\"10666339\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2001,\n      \"finding\": \"AFAP-110(Δlzip), a leucine zipper deletion mutant, activates cellular tyrosine kinases including Src family members and becomes hyperphosphorylated on tyrosine. This activation requires integrity of the SH3-binding motif of AFAP-110(Δlzip). A deletion within the PH domain also reverts the effects on actin filaments. Dominant-positive RhoA(V14) blocks AFAP-110(Δlzip)-induced actin filament rosettes but not Src activation, placing Rho downstream of AFAP-110-directed Src activation in the actin reorganization pathway.\",\n      \"method\": \"Expression of deletion and point mutants, immunofluorescence, kinase activity assays, co-expression with dominant-active RhoA\",\n      \"journal\": \"Oncogene\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — epistasis via dominant-active RhoA + mutagenesis + cellular phenotype, single lab multiple methods\",\n      \"pmids\": [\"11641786\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2002,\n      \"finding\": \"Recombinant AFAP-110 binds actin filaments cooperatively through lateral association and can cross-link actin filaments in a manner dependent on the carboxy-terminal actin-binding domain. Deletion of the leucine zipper or PKC phosphorylation alters AFAP-110 conformation, reduces multimerization stability, and increases its capacity to cross-link actin filaments. PKC activation in cells causes loss of stress fibers, formation of motility structures, and strong AFAP-110 colocalization with these structures. Expression of an AFAP-110 mutant unable to bind PKC blocked PMA effects on actin filaments.\",\n      \"method\": \"In vitro actin cross-linking assay with recombinant AFAP-110, PKC phosphorylation in vitro, Superose chromatography, expression of PKC-binding domain deletion mutant, phorbol ester treatment and immunofluorescence\",\n      \"journal\": \"Molecular biology of the cell\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — in vitro reconstitution of actin cross-linking + phosphorylation + mutagenesis + cellular imaging, single lab, multiple orthogonal methods\",\n      \"pmids\": [\"12134071\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 1998,\n      \"finding\": \"AFAP-110 forms a stable complex with activated Src through either of two independently functional SH2-binding motifs: one in the amino terminus (Y94) and one in the carboxy terminus (Y451 and Y453). Mutation of Y93/Y94 or Y451/Y453 alone does not fully abrogate complex formation, but triple-tyrosine mutations (combining both regions) drastically reduce phosphorylation and prevent stable complex formation. A multistep SH2/SH3 binding mechanism is supported.\",\n      \"method\": \"Site-directed mutagenesis of candidate SH2-binding tyrosines, co-expression with Src527F in Cos-1 cells, co-immunoprecipitation, tyrosine phosphorylation analysis\",\n      \"journal\": \"Molecular carcinogenesis\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — systematic site-directed mutagenesis combined with co-IP and phosphorylation assays, single lab multiple complementary approaches\",\n      \"pmids\": [\"9655255\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2004,\n      \"finding\": \"The leucine zipper motif of AFAP-110 regulates actin filament cross-linking and Src activation through both sequence and structural requirements. The leucine zipper facilitates intramolecular auto-inhibition by contacting the amino-terminal PH1 domain. Disruption of multimer stability (by Δlzip, fos-Lzip substitution, or L581P helix mutation) increases in vitro actin cross-linking; however, only structural disruptions (Δlzip, L581P) activate cSrc and alter actin integrity in vivo, indicating that actin cross-linking and Src activation are distinct functions.\",\n      \"method\": \"Leucine zipper substitution and point mutagenesis, in vitro actin cross-linking assay, cellular expression with immunofluorescence and cSrc activation assays, opposing binding site analysis\",\n      \"journal\": \"Journal of cellular biochemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — in vitro reconstitution + multiple mutagenesis strategies + cellular phenotype, single lab with clear mechanistic dissection\",\n      \"pmids\": [\"14755689\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2007,\n      \"finding\": \"AFAP-110 (AFAP1) is required for actin stress fiber formation and cell adhesion to fibronectin in MDA-MB-231 breast cancer cells. Knockdown of AFAP-110 abolishes stress fiber cross-linking and focal adhesion formation without affecting proliferation or expression of focal adhesion components. LPA treatment cannot rescue stress fiber and focal adhesion formation in AFAP-110 knockdown cells.\",\n      \"method\": \"siRNA knockdown, inducible knockdown, immunofluorescence, fibronectin adhesion assay, LPA stimulation\",\n      \"journal\": \"Journal of cellular physiology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — clean RNAi knockdown with inducible confirmation + defined cellular phenotype (stress fibers, focal adhesions), single lab two orthogonal methods\",\n      \"pmids\": [\"17520695\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2007,\n      \"finding\": \"AFAP-110 (AFAP1) is required for tumorigenic growth of prostate cancer cells, regulating focal contacts in a PKC-dependent manner. Downregulation of AFAP-110 reduced cell-matrix adhesion, migration, defective focal adhesions, and reduced integrin β1 expression. Reintroduction of avian AFAP-110 or a Src-interaction-disabled mutant restored these properties, but an AFAP-110 lacking the PKC-interacting domain failed to restore them, demonstrating PKC-dependent regulation of focal contacts.\",\n      \"method\": \"Immunohistochemistry on human tissue arrays, siRNA knockdown, orthotopic nude mouse model, reintroduction of AFAP-110 domain mutants, cell adhesion and migration assays, integrin β1 expression analysis\",\n      \"journal\": \"The Journal of clinical investigation\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — domain mutant rescue experiments + in vivo orthotopic model + multiple phenotypic readouts, single rigorous study\",\n      \"pmids\": [\"17885682\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2007,\n      \"finding\": \"PI3K activation is required for PMA/PKCα-induced colocalization of AFAP-110 with c-Src and subsequent c-Src activation. The amino-terminal PH1 domain of AFAP-110 is required for colocalization with c-Src, while the SH3-binding motif is required for activation. PI3K inhibitors and cells lacking p85α/β PI3K regulatory subunits block AFAP-110–c-Src colocalization and activation. This pathway promotes cell migration.\",\n      \"method\": \"PI3K inhibitor treatment, PI3K p85 knockout cells, dominant-negative AFAP-110 expression, wound healing migration assay, immunofluorescence, domain mutant analysis\",\n      \"journal\": \"American journal of physiology. Cell physiology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — genetic (knockout) and pharmacological PI3K inhibition + domain mutant rescue + migration assay, single lab multiple orthogonal approaches\",\n      \"pmids\": [\"17360811\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2008,\n      \"finding\": \"PKCα phosphorylates AFAP-110 at Ser277 in response to phorbol ester treatment, and phospho-Ser277 AFAP-110 localizes to podosomes. Mutation of Ser277 to Ala (AFAP-110-S277A) does not alter podosome localization but increases podosome number and lifespan (demonstrated by video microscopy), indicating that Ser277 phosphorylation regulates podosome stability and turnover.\",\n      \"method\": \"Mutational analysis, phospho-specific antibody generation and immunolabeling, video microscopy of podosome dynamics in A7r5 vascular smooth muscle cells\",\n      \"journal\": \"Journal of cell science\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — phospho-specific antibody validation + mutagenesis + live-cell video microscopy, single lab multiple methods\",\n      \"pmids\": [\"18577577\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2009,\n      \"finding\": \"Tks5 recruits AFAP-110, p190RhoGAP, and cortactin to podosomes via its fifth SH3 domain. Mislocalization of Tks5 to mitochondria causes redistribution of AFAP-110, p190RhoGAP, and cortactin to perinuclear regions and inhibits podosome formation. A Tks5 mutant lacking the PX domain also inhibits podosome formation and redistributes AFAP-110.\",\n      \"method\": \"Forced mitochondrial mislocalization of Tks5 deletion mutants, immunofluorescence, siRNA knockdown of Tks5 and p190RhoGAP\",\n      \"journal\": \"Experimental cell research\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — subcellular mislocalization experiment + domain deletion mutant series + siRNA knockdown, single lab multiple approaches\",\n      \"pmids\": [\"19540230\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 1995,\n      \"finding\": \"An alternatively spliced brain-specific variant of AFAP-110, designated AFAP-120, contains an additional 258 bp open reading frame encoding a novel internal sequence (NINS) with a proline-rich SH3-binding motif. The NINS-GST fusion protein pulls down a 67-kDa protein from tissue lysates; deletion analysis maps this interaction to the proline-rich motif.\",\n      \"method\": \"cDNA cloning, northern blot, western blot, GST-NINS pulldown assay with deletion mutants, transient expression\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 / Moderate — single pulldown experiment identifying novel binding partner, corroborated by deletion analysis, single lab\",\n      \"pmids\": [\"7876134\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2011,\n      \"finding\": \"Endogenous AFAP1 localizes to both cytoskeletal and cytosolic fractions; depolymerization of actin filaments (cytochalasin D or latrunculin A) increases AFAP1 in the cytosolic fraction, demonstrating actin-dependent localization. An AFAP1 mutant lacking the actin-binding domain (AFAPΔABD) localizes predominantly to the cytosolic fraction, is associated with disorganized cytoskeleton, reduced stress fibers, and accumulation of F-actin at the membrane and podosome-like structures that co-localize with cortactin but do not digest gelatin, and show reduced interaction with c-Src.\",\n      \"method\": \"Subcellular fractionation, cytoskeletal drug treatment, overexpression of ABD deletion mutant, immunofluorescence, gelatin degradation assay\",\n      \"journal\": \"Cellular and molecular life sciences : CMLS\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — fractionation with functional drug perturbations + mutagenesis + multiple cellular phenotype readouts, single lab with multiple orthogonal methods\",\n      \"pmids\": [\"21984596\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2011,\n      \"finding\": \"AFAP1L1, a paralog of AFAP1 in the AFAP family, preferentially interacts with the SH3 domain of cortactin (not c-Src), in contrast to AFAP1 which binds cSrc SH3 but not cortactin. Overexpression of AFAP1L1 in A7r5 cells induces podosome formation without stimulation and colocalizes with cortactin at invadosomes.\",\n      \"method\": \"SH3 domain binding assay, fluorescence microscopy, overexpression in A7r5 cells, immunohistochemistry\",\n      \"journal\": \"European journal of cell biology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 / Moderate — SH3 binding specificity comparison + cellular overexpression phenotype; this is primarily about AFAP1L1 but establishes AFAP1's selectivity for cSrc SH3 over cortactin by direct comparison\",\n      \"pmids\": [\"21333378\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"AFAP1 (the protein) is a key mediator of TNF-α-induced rapid attenuation of P-glycoprotein (P-gp) efflux activity at the blood-brain barrier. TNF-α rapidly decreases P-gp efflux activity without changing P-gp protein expression. Phosphoproteomics identified AFAP1 as significantly phosphorylated upon TNF-α treatment. Knockdown of AFAP1 blocked the TNF-α-induced reduction in P-gp efflux activity, while MAPK1 inhibition or AP-1 knockdown did not, demonstrating a specific non-transcriptional role for AFAP1 in inflammatory modulation of P-gp function.\",\n      \"method\": \"Quantitative phosphoproteomics, siRNA knockdown of AFAP1, P-gp efflux activity assay, western blot for P-gp expression in plasma membrane fraction, in vivo LPS mouse model\",\n      \"journal\": \"Journal of neurochemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — quantitative phosphoproteomics + selective siRNA knockdown with functional readout + in vivo validation + multiple negative controls (MAPK1, AP-1), single rigorous study\",\n      \"pmids\": [\"28112407\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"AFAP1 (AFAP-110) is an adaptor and actin cross-linking protein that integrates PKCα and PI3K signals to activate c-Src and reorganize the actin cytoskeleton: PKCα phosphorylates AFAP1 at Ser277, PI3K activity enables its PH1 domain-dependent colocalization with c-Src, and its SH3-binding motif engages the c-Src SH3 domain to drive kinase activation, podosome formation, focal adhesion regulation, and cell migration; these activities are regulated by an intramolecular auto-inhibitory mechanism in which the carboxy-terminal leucine zipper constrains multimerization and actin cross-linking, and by Src-induced conformational changes that relieve this inhibition; additionally, AFAP1 mediates inflammatory (TNF-α/PKC) attenuation of P-glycoprotein efflux at the blood-brain barrier through a non-transcriptional phosphorylation-dependent mechanism.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"AFAP1 (AFAP-110) is a multidomain adaptor and actin cross-linking protein that couples PKC\\u03b1 and PI3K signaling to c-Src activation and actin cytoskeletal remodeling, driving podosome formation, focal adhesion regulation, and cell migration [#0, #11]. Its amino terminus carries SH3- and SH2-binding modules that engage activated Src: an SH3-binding motif mediates stable complex formation and efficient tyrosine phosphorylation, while two independently functional SH2-binding regions (Y94 and Y451/Y453) cooperate in a multistep binding mechanism [#2, #7]. The carboxy-terminal actin-binding domain is necessary and sufficient for direct, cooperative actin filament binding and cross-linking, and is required for cytoskeletal localization [#4, #6, #15]. These activities are held under intramolecular auto-inhibition by a carboxy-terminal leucine zipper that contacts the PH1 domain and constrains multimerization; structural disruption of the zipper, or activated-Src-induced conformational change, relieves this inhibition and separates the distinct outputs of actin cross-linking versus Src activation [#3, #8]. Upstream, PKC\\u03b1 phosphorylates AFAP1 at Ser277 to regulate podosome stability and turnover, PI3K activity and the PH1 domain enable AFAP1\\u2013c-Src colocalization, and Rho acts downstream in the actin reorganization arm [#5, #11, #12]. AFAP1 is required for stress fiber and focal adhesion formation, integrin \\u03b21-dependent adhesion, and tumorigenic growth of prostate and breast cancer cells [#9, #10], and it is recruited to podosomes by the Tks5 fifth SH3 domain [#13]. Independently, AFAP1 is a phosphorylation-dependent, non-transcriptional mediator of TNF-\\u03b1\\u2013induced attenuation of P-glycoprotein efflux at the blood-brain barrier [#17].\",\n  \"teleology\": [\n    {\n      \"year\": 1995,\n      \"claim\": \"Identifying a brain-specific alternatively spliced variant established that the AFAP locus encodes multiple proteins and revealed protein-interaction modules through a novel internal proline-rich SH3-binding sequence.\",\n      \"evidence\": \"cDNA cloning, northern/western blot, and GST-NINS pulldown of a 67-kDa partner in tissue lysates\",\n      \"pmids\": [\"7876134\"],\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"\",\n      \"gaps\": [\"Identity of the 67-kDa interacting protein not determined\", \"Functional role of the AFAP-120 variant not established\"]\n    },\n    {\n      \"year\": 1997,\n      \"claim\": \"Mapping the amino-terminal SH3-binding motif answered how AFAP-110 docks onto Src, showing this motif is required for stable Src complex formation and efficient tyrosine phosphorylation.\",\n      \"evidence\": \"Site-directed mutagenesis, GST-SH3 pulldown, and co-IP with activated Src527F in Cos-1 cells\",\n      \"pmids\": [\"9350057\"],\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"\",\n      \"gaps\": [\"Did not address the contribution of SH2-binding tyrosines to complex formation\", \"Consequences for downstream actin remodeling not tested\"]\n    },\n    {\n      \"year\": 1998,\n      \"claim\": \"Defining two SH2-binding tyrosine regions and a self-associating leucine zipper resolved the architecture of Src binding and revealed activated Src remodels AFAP-110 into multimers.\",\n      \"evidence\": \"Tyrosine mutagenesis with co-IP/phosphorylation analysis, plus GST affinity absorption, size-exclusion chromatography, and deletional mutagenesis with imaging in Src527F cells\",\n      \"pmids\": [\"9655255\", \"9619827\"],\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"\",\n      \"gaps\": [\"Mechanism coupling multimer state to Src activation not yet defined\", \"In vitro reconstitution of the conformational switch not performed\"]\n    },\n    {\n      \"year\": 2000,\n      \"claim\": \"Localizing direct actin-filament binding to the carboxy terminus, and showing leucine zipper deletion unleashes actin-altering activity, established that the leucine zipper auto-inhibits AFAP-110's intrinsic cytoskeletal function.\",\n      \"evidence\": \"In vitro actin binding with recombinant C-terminal domain, deletional mutagenesis, and immunofluorescence in untransformed fibroblasts\",\n      \"pmids\": [\"10666339\"],\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"\",\n      \"gaps\": [\"Whether actin alteration requires kinase activation not resolved here\", \"Structural basis of auto-inhibition not defined\"]\n    },\n    {\n      \"year\": 2001,\n      \"claim\": \"Epistasis experiments placed AFAP-110-directed Src activation upstream of Rho-dependent actin reorganization and showed the SH3-binding motif and PH domain are required for these activities.\",\n      \"evidence\": \"Expression of \\u0394lzip and point mutants, kinase assays, and co-expression with dominant-active RhoA(V14)\",\n      \"pmids\": [\"11641786\"],\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"\",\n      \"gaps\": [\"Direct Rho activation mechanism by AFAP-110 not shown\", \"Physiological trigger for the conformational change not identified\"]\n    },\n    {\n      \"year\": 2002,\n      \"claim\": \"Reconstituting cooperative actin cross-linking and showing PKC phosphorylation alters AFAP-110 conformation linked an upstream kinase to its cytoskeletal cross-linking function.\",\n      \"evidence\": \"In vitro actin cross-linking and PKC phosphorylation of recombinant AFAP-110, size-exclusion chromatography, and PMA treatment with a PKC-binding-deficient mutant\",\n      \"pmids\": [\"12134071\"],\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"\",\n      \"gaps\": [\"Specific PKC phosphorylation site not yet mapped\", \"Quantitative relationship between multimer destabilization and cross-linking not defined\"]\n    },\n    {\n      \"year\": 2004,\n      \"claim\": \"Two studies established that PKC\\u03b1 signaling requires AFAP-110 to activate c-Src and form podosomes, and dissected the leucine zipper into separable sequence (cross-linking) and structural (Src activation) determinants contacting the PH1 domain.\",\n      \"evidence\": \"Rescue with domain mutants in AFAP-110-deficient CaOV3 cells, plus leucine zipper substitution/point mutagenesis with in vitro cross-linking and cellular cSrc activation assays\",\n      \"pmids\": [\"15314167\", \"14755689\"],\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"\",\n      \"gaps\": [\"Atomic structure of the PH1\\u2013leucine zipper auto-inhibitory contact not solved\", \"How PKC\\u03b1 binding triggers the conformational relief not detailed\"]\n    },\n    {\n      \"year\": 2007,\n      \"claim\": \"Knockdown and in vivo tumor studies established AFAP1 as required for stress fiber and focal adhesion formation, integrin \\u03b21-dependent adhesion, migration, and tumorigenic growth, with PI3K positioned upstream of AFAP1\\u2013c-Src colocalization.\",\n      \"evidence\": \"siRNA/inducible knockdown with adhesion and imaging assays in breast cancer cells; domain-mutant rescue plus orthotopic mouse model in prostate cancer cells; PI3K inhibition/p85 knockout with wound-healing assay\",\n      \"pmids\": [\"17520695\", \"17885682\", \"17360811\"],\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"\",\n      \"gaps\": [\"Direct link between PI3K lipid products and the PH1 domain not biochemically demonstrated\", \"How AFAP1 loss reduces integrin \\u03b21 expression not mechanistically explained\"]\n    },\n    {\n      \"year\": 2008,\n      \"claim\": \"Identifying Ser277 as the PKC\\u03b1 phosphorylation site and showing the S277A mutant increases podosome number and lifespan defined a regulatory phosphorylation controlling podosome turnover.\",\n      \"evidence\": \"Phospho-specific antibody, mutagenesis, and live-cell video microscopy of podosome dynamics in A7r5 cells\",\n      \"pmids\": [\"18577577\"],\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"\",\n      \"gaps\": [\"Downstream effectors reading the Ser277 phosphorylation state not identified\", \"Relationship of Ser277 to the leucine zipper/PH1 auto-inhibition not tested\"]\n    },\n    {\n      \"year\": 2009,\n      \"claim\": \"Mislocalization experiments showed Tks5 recruits AFAP-110 (with p190RhoGAP and cortactin) to podosomes via its fifth SH3 domain, defining a scaffold that positions AFAP1 at invadosomal structures.\",\n      \"evidence\": \"Forced mitochondrial mislocalization of Tks5 deletion mutants, immunofluorescence, and siRNA knockdown\",\n      \"pmids\": [\"19540230\"],\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"\",\n      \"gaps\": [\"Whether Tks5\\u2013AFAP1 binding is direct not established\", \"Hierarchy of recruitment among AFAP1, cortactin and p190RhoGAP unresolved\"]\n    },\n    {\n      \"year\": 2011,\n      \"claim\": \"Fractionation, ABD deletion, and paralog comparison clarified that actin-bound localization governs AFAP1 cytoskeletal function and Src engagement, and that AFAP1 selectively binds c-Src SH3 (versus cortactin) distinguishing it from AFAP1L1.\",\n      \"evidence\": \"Subcellular fractionation with actin-depolymerizing drugs and ABD-deletion imaging/gelatin assay; SH3-domain binding specificity comparison and overexpression in A7r5 cells\",\n      \"pmids\": [\"21984596\", \"21333378\"],\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"\",\n      \"gaps\": [\"Functional consequence of AFAP1 selectivity for c-Src over cortactin in vivo not tested\", \"Whether ABD-dependent localization is regulated by signaling not addressed\"]\n    },\n    {\n      \"year\": 2017,\n      \"claim\": \"Phosphoproteomics and selective knockdown identified AFAP1 as a non-transcriptional mediator of TNF-\\u03b1\\u2013induced attenuation of P-glycoprotein efflux at the blood-brain barrier, extending AFAP1 function beyond cytoskeletal/Src signaling.\",\n      \"evidence\": \"Quantitative phosphoproteomics, siRNA knockdown with P-gp efflux assay, negative controls (MAPK1, AP-1), and in vivo LPS mouse model\",\n      \"pmids\": [\"28112407\"],\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"\",\n      \"gaps\": [\"Phosphorylation site(s) on AFAP1 driving P-gp modulation not mapped\", \"Mechanism by which phospho-AFAP1 reduces P-gp transport activity unknown\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"How AFAP1's well-defined cytoskeletal/Src adaptor activities mechanistically connect to its non-transcriptional regulation of P-glycoprotein at the blood-brain barrier remains unresolved.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"\",\n      \"gaps\": [\"No structure of full-length AFAP1 or its auto-inhibited state\", \"The effector linking phospho-AFAP1 to P-gp transport activity is unidentified\", \"Direct biochemical link between PI3K products and the PH1 domain not demonstrated\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0060090\", \"supporting_discovery_ids\": [0, 1, 11]},\n      {\"term_id\": \"GO:0008092\", \"supporting_discovery_ids\": [4, 6, 15]},\n      {\"term_id\": \"GO:0005198\", \"supporting_discovery_ids\": [6, 9]},\n      {\"term_id\": \"GO:0098772\", \"supporting_discovery_ids\": [0, 5, 8]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005856\", \"supporting_discovery_ids\": [4, 6, 15]},\n      {\"term_id\": \"GO:0005829\", \"supporting_discovery_ids\": [15]},\n      {\"term_id\": \"GO:0005886\", \"supporting_discovery_ids\": [3, 15]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-162582\", \"supporting_discovery_ids\": [0, 5, 11]},\n      {\"term_id\": \"R-HSA-1474244\", \"supporting_discovery_ids\": [9, 10]},\n      {\"term_id\": \"R-HSA-1643685\", \"supporting_discovery_ids\": [10, 17]}\n    ],\n    \"complexes\": [\"podosome\", \"focal adhesion\"],\n    \"partners\": [\"SRC\", \"PRKCA\", \"TKS5\", \"CTTN\", \"PIK3R1\"],\n    \"other_free_text\": []\n  }\n}","audit_flag":null,"evaluation":{"pairwise":"win","faith_supported":7,"faith_total":7,"faith_pct":100.0}}