{"gene":"EPHB1","run_date":"2026-06-09T23:54:43","timeline":{"discoveries":[{"year":1999,"finding":"EphB1 functions as a 'ligand density sensor': engagement with ephrin-B1 at high surface density activates integrin-mediated cell attachment (alphavbeta3 in endothelial cells; alpha5beta1 in transfected HEK cells) without mechanical tethering. Activation-competent but signaling-defective EphB1 point mutants failed to stimulate ephrin-B1-dependent attachment, demonstrating requirement for EphB1 kinase signaling.","method":"Reconstituted ephrin-B1 surface density assay, integrin-blocking antibodies, EphB1 point mutants in transfection experiments, cell attachment assays","journal":"The EMBO journal","confidence":"High","confidence_rationale":"Tier 1 / Moderate — in vitro reconstitution with point-mutant validation, single lab but multiple orthogonal methods (density assay + integrin blocking + mutagenesis)","pmids":["10205170"],"is_preprint":false},{"year":1998,"finding":"The adapter protein Nck binds directly to the juxtamembrane region of EphB1 via its SH2 domain, requiring phospho-Tyr594. Ligand (ephrin-B1/Fc) activation of EphB1 recruits Nck to native receptor complexes and activates c-Jun kinase (JNK/SAPK). Mutant EphB1-Y594F blocks Nck recruitment, attenuates JNK activation, and blocks cell attachment responses.","method":"Yeast two-hybrid cloning, Co-immunoprecipitation, site-directed mutagenesis (Y594F), JNK kinase assay, cell attachment assay","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 1-2 / Moderate — yeast two-hybrid + Co-IP + mutagenesis + kinase assay in single lab with multiple orthogonal methods","pmids":["9430661"],"is_preprint":false},{"year":2003,"finding":"Activated EphB1 recruits adaptor proteins Grb2 and p52Shc, promotes c-Src-dependent tyrosine phosphorylation of p52Shc, and activates MAPK/ERK to drive cell chemotaxis. EphB1 Tyr600 and Tyr778 are required for interaction with c-Src and p52Shc. Phosphorylation of p52Shc by c-Src is required for its recruitment to EphB1 complexes via its phosphotyrosine binding domain. ERK and Src inhibitors abolished EphB1-mediated migration but not adhesion.","method":"Co-immunoprecipitation, site-directed mutagenesis (Y600F, Y778F), dominant-negative c-Src expression, pharmacological inhibitors (PD98059, PP2), cell migration and adhesion assays","journal":"The Journal of cell biology","confidence":"High","confidence_rationale":"Tier 1-2 / Moderate — reciprocal Co-IP, mutagenesis, dominant-negative, and pharmacological approaches in single lab","pmids":["12925710"],"is_preprint":false},{"year":2002,"finding":"Grb7 binds to the cytoplasmic domain of EphB1 via its SH2 domain in a manner requiring EphB1 autophosphorylation. Tyr-928 of EphB1 is the primary Grb7 binding site. EphB1 can phosphorylate Grb7, and mutations Y928F or Y594F reduced this activity. Co-expression of Grb7 with EphB1 enhanced cell motility, while the Grb7 SH2 domain alone abolished EphB1-stimulated migration.","method":"Yeast two-hybrid screening, Co-immunoprecipitation, site-directed mutagenesis, kinase assay, cell migration assay on extracellular matrix","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 2 / Moderate — yeast two-hybrid + Co-IP + mutagenesis + functional migration assay in single lab","pmids":["12223469"],"is_preprint":false},{"year":2001,"finding":"The kinase-inactive EphB6 receptor undergoes transphosphorylation upon ligand (ephrin-B1) stimulation when co-expressed with catalytically active EphB1. EphB1 and EphB6 form a stable hetero-complex; EphB1-induced EphB6 phosphorylation requires EphB1 catalytic activity. The proto-oncogene c-Cbl was identified as a constitutive EphB6-binding protein requiring a functional phosphotyrosine binding domain.","method":"Co-immunoprecipitation, overexpression of kinase-dead EphB1 mutants, ligand stimulation assays","journal":"The Journal of biological chemistry","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — reciprocal Co-IP with mutant controls, single lab, two methods","pmids":["11713248"],"is_preprint":false},{"year":2004,"finding":"Activated EphB1 induces c-Src-dependent tyrosine phosphorylation of paxillin at Tyr-31 and Tyr-118, and is recruited to paxillin-FAK complexes. Cells expressing paxillin Y31F/Y118F or paxillin ΔLD4 show reduced EphB1-dependent migration. Nck binding site mutation (Y594F) disrupts the Nck-paxillin-EphB1 complex, establishing a signaling complex: EphB1 → Nck → paxillin/FAK → c-Src-mediated paxillin phosphorylation → cell migration.","method":"Co-immunoprecipitation, site-directed mutagenesis (paxillin Y31F/Y118F, EphB1 Y594F), dominant-negative c-Src, Src inhibitor PP2, cell migration assay","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 1-2 / Moderate — mutagenesis of both receptor and substrate, Co-IP, dominant-negative, inhibitor validation in single lab","pmids":["15107421"],"is_preprint":false},{"year":2006,"finding":"EphB1 receptors localize to caveolae and directly interact with caveolin-1 (Cav-1) upon ligand stimulation. This interaction requires the caveolin-binding motif within the EphB1 kinase domain, which is also required for correct membrane targeting of EphB1. Overexpression of a Cav-1 mutant lacking the scaffolding domain abolishes EphB1-Cav-1 interaction and EphB1-mediated ERK activation.","method":"Subcellular fractionation, co-immunoprecipitation, caveolin-1 scaffolding domain mutant overexpression, confocal microscopy, ERK activation assays","journal":"Journal of cell science","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — Co-IP, fractionation, dominant-negative mutant, and signaling assay in single lab","pmids":["16723736"],"is_preprint":false},{"year":2007,"finding":"Ligand (ephrin-B1) stimulation of EphB1 leads to Cbl recruitment to EphB1 via Cbl's tyrosine kinase-binding domain, followed by Src-dependent Cbl phosphorylation, EphB1 ubiquitination, and lysosomal degradation of EphB1. Overexpression of wild-type but not the ligase-dead 70Z Cbl mutant enhanced EphB1 ubiquitination and degradation. Kinase-dead EphB1-K652R was resistant to Cbl-mediated degradation.","method":"Co-immunoprecipitation, GST pull-down, overexpression of Cbl mutants, ubiquitination assay, lysosomal inhibitor (bafilomycin) treatment, Src inhibitor (PP2)","journal":"Traffic (Copenhagen, Denmark)","confidence":"High","confidence_rationale":"Tier 1-2 / Moderate — GST pull-down + Co-IP + ubiquitination assay + multiple mutants in single lab","pmids":["18034775"],"is_preprint":false},{"year":2003,"finding":"Ephrin-B2 is expressed at the optic chiasm midline and selectively repels ventrotemporal retinal ganglion cell (RGC) axons that express EphB1. EphB1 is found exclusively in retinal regions giving rise to the ipsilateral projection. EphB1 null mice exhibit a dramatically reduced ipsilateral retinal projection, and blocking ephrin-B2 function in vitro eliminates the inhibitory effect on chiasm cells and abolishes ipsilateral projection in semiintact preparation.","method":"Immunohistochemistry, EphB1 knockout mouse, in vitro axon growth assays with function-blocking reagents, semiintact visual system preparation","journal":"Neuron","confidence":"High","confidence_rationale":"Tier 2 / Strong — genetic knockout + in vitro blocking assays + expression localization, replicated in subsequent studies","pmids":["12971893"],"is_preprint":false},{"year":1997,"finding":"EphB1 (and EphA4) expressed in migrating third arch neural crest cells interact with ephrin-B2 in second arch neural crest/mesoderm to restrict intermingling and direct targeted migration. Inhibition of EphB1/EphA4 function using truncated receptors leads to abnormal third arch neural crest migration into second and fourth arch territories; ectopic ephrin-B2 overexpression scatters third arch cells.","method":"Dominant-negative truncated receptor expression in Xenopus embryos, ectopic ephrin-B2 overexpression, in situ hybridization, cell lineage tracing","journal":"Current biology : CB","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — dominant-negative and gain-of-function in Xenopus embryos, single lab","pmids":["9259557"],"is_preprint":false},{"year":1999,"finding":"EphB1 and its ligand ephrin-B2 are expressed in complementary patterns in midbrain dopaminergic neurons and their targets. Ephrin-B2 selectively inhibits neurite outgrowth and induces cell loss of substantia nigra (but not ventral tegmental) dopaminergic neurons, suggesting EphB1/ephrin-B2 specifies distinct dopaminergic pathways. Ephrin-B2 expression is upregulated by cocaine and amphetamine in adults.","method":"Neurite outgrowth inhibition assay, cell survival assay, in situ hybridization, immunohistochemistry","journal":"The Journal of neuroscience","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — in vitro functional assays with specific ligand, expression pattern analysis, single lab","pmids":["10066262"],"is_preprint":false},{"year":2008,"finding":"Zic2 transcription factor regulates EphB1 expression in retinal ganglion cells. Ectopic delivery of Zic2 into non-VT retinal explants induces EphB1 mRNA and protein expression. The upregulated EphB1 localizes to growth cones and is functional, converting RGC axon behavior from extension onto to avoidance of ephrin-B2 substrates, demonstrating Zic2→EphB1 axis controls ipsilateral projection.","method":"Retinal explant electroporation, in vitro axon growth/avoidance assays on ephrin-B2 substrates, immunofluorescence for EphB1 protein in growth cones","journal":"The Journal of neuroscience","confidence":"High","confidence_rationale":"Tier 2 / Moderate — gain-of-function with functional readout, protein localization, multiple assays in single lab","pmids":["18524895"],"is_preprint":false},{"year":2009,"finding":"EphB1 is specifically required and sufficient to drive the ipsilateral retinal projection. In utero retinal electroporation of EphB1 redirects normally-crossed RGCs to ipsilateral trajectory. EphB2, despite high similarity, is far less efficient. EphB1-EphB2 chimeric receptor analysis reveals that both extracellular and juxtamembrane domains of EphB1 are specifically required for ipsilateral redirection.","method":"In utero retinal electroporation, chimeric receptor expression, in vivo axon tracing","journal":"The Journal of neuroscience","confidence":"High","confidence_rationale":"Tier 2 / Moderate — in vivo gain-of-function with chimeric receptor dissection, domain-specific analysis","pmids":["19295152"],"is_preprint":false},{"year":2011,"finding":"EphB1 forward signaling (into the EphB1-expressing cell) is specifically required for ipsilateral retinal axon projection; reverse signaling through the EphB1 extracellular domain is not required. Knock-in mice expressing intracellular-truncated EphB1 (EphB1T-lacZ) fail to form the ipsilateral projection, phenocopying the EphB1 null. EphB1 is the preferred receptor for ephrin-B2 (and less so ephrin-B1) at the optic chiasm.","method":"Knock-in mouse (intracellular truncation), retinal axon tracing, genetic analysis of EphB/ephrin-B mutant combinations","journal":"The European journal of neuroscience","confidence":"High","confidence_rationale":"Tier 2 / Moderate — clean knock-in distinguishing forward vs. reverse signaling, in vivo tracing","pmids":["22103419"],"is_preprint":false},{"year":2008,"finding":"EphB1 null mice show a 40% decrease in neuron number and volume in substantia nigra pars reticulata (SNr), but not in pars compacta TH+ neurons. EphB1 is expressed in SNr (not SNc) neurons. Loss of EphB1 results in spontaneous locomotor hyperactivity, linking EphB1-dependent SNr development to locomotor function.","method":"EphB1 knockout mice, stereological neuron counting, in situ hybridization, beta-galactosidase reporter, open-field behavioral testing","journal":"The European journal of neuroscience","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — genetic knockout with anatomical and behavioral readout, single lab","pmids":["17561836"],"is_preprint":false},{"year":2012,"finding":"EphB1 recruits scaffold protein Caskin1 via the adaptor Nck: EphB1 activation leads to Nck SH2 binding to phosphotyrosine on EphB1, while Nck SH3 domains interact with the proline-rich domain of Caskin1. This complex formation results in tyrosine phosphorylation of Caskin1 at Tyr296 and Tyr336 (identified by mass spectrometry), and phosphorylation causes structural changes in the Caskin1 SH3 domain detected by CD spectroscopy.","method":"Co-immunoprecipitation, mass spectrometry phosphosite identification, CD spectroscopy","journal":"Cell communication and signaling : CCS","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — Co-IP + MS phosphosite + structural CD spectroscopy in single lab","pmids":["23181695"],"is_preprint":false},{"year":2012,"finding":"PTEN is constitutively associated with c-Cbl, protecting Cbl from degradation. EphB1 stimulation triggers PTEN dephosphorylation (~50% on Ser/Thr) and disruption of the PTEN-Cbl complex, requiring PTEN protein phosphatase activity. Both PTEN and Cbl independently translocate to EphB1, with PTEN associating via scaffold protein NHERF1. PTEN lipid phosphatase activity impairs EphB1-dependent cell adhesion and chemotaxis.","method":"Co-immunoprecipitation, PTEN and NHERF1 siRNA knockdown, PTEN phosphatase mutants, cell adhesion and chemotaxis assays","journal":"FASEB journal","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — Co-IP, siRNA, and phosphatase-dead mutants with functional assays, single lab","pmids":["23118026"],"is_preprint":false},{"year":2017,"finding":"EphB1 is upregulated in injured motor neurons and can activate astrocytes through ephrin-B1-mediated stimulation of STAT3. EphB1-induced astrocyte activation produces a protective and anti-inflammatory transcriptional signature distinct from IL-6-induced responses. This EphB1-ephrin-B1-STAT3 pathway is disrupted in human stem cell-derived astrocytes and mouse models of ALS.","method":"Transcriptional analysis of EphB1-stimulated astrocytes, STAT3 pathway analysis, human iPSC-derived astrocytes, ALS mouse models","journal":"Nature communications","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — multiple model systems (human iPSC and mouse), pathway analysis, single lab","pmids":["29079839"],"is_preprint":false},{"year":2018,"finding":"The RhoGAP protein Porf-2 is an intracellular mediator of EphB1 signaling in axon growth inhibition. EphB1 binds and regulates Porf-2 upon ephrin-B stimulation. The activated EphB1 forward signal deactivates Rac1 through the GAP domain of Porf-2, which inhibits growth cone expansion and brakes axon outgrowth.","method":"Co-immunoprecipitation of EphB1-Porf-2, Rac1 activity assays (GTP-Rac1 pull-down), axon growth measurement, GAP domain mutagenesis","journal":"Cellular and molecular life sciences : CMLS","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — Co-IP, Rac1 activity assay, functional axon growth readout in single lab","pmids":["29938386"],"is_preprint":false},{"year":2020,"finding":"EphB1 constitutively interacts with caveolin-1 (Cav-1) in endothelial cells. Ephrin-B1 activation of EphB1 induces EphB1 phosphorylation, uncouples EphB1 from Cav-1, and promotes Src-dependent Cav-1 phosphorylation. Deletion of the CSD-binding motif in EphB1 prevents Cav-1 interaction and Cav-1 phosphorylation. EphB1-/- ECs show markedly reduced Cav-1 expression and caveolae numbers due to Cav-1 ubiquitination and degradation when not bound to EphB1.","method":"Super-resolution microscopy, FRET, co-immunoprecipitation, EphB1 CSD-motif deletion mutant, EphB1 knockout endothelial cells, electron microscopy caveolae counting","journal":"Molecular biology of the cell","confidence":"High","confidence_rationale":"Tier 1-2 / Strong — FRET + Co-IP + mutagenesis + knockout ECs + structural imaging, multiple orthogonal methods","pmids":["32238105"],"is_preprint":false},{"year":2021,"finding":"Tetracycline antibiotics demeclocycline, chlortetracycline, and minocycline inhibit EphB1 kinase activity at low micromolar concentrations by binding to the ATP-binding catalytic domain. Co-crystallization of chlortetracycline with EphB1 confirmed binding to the ATP-binding domain. In vivo, the three-tetracycline combination inhibited EphB1 phosphorylation in brain, spinal cord, and DRG and blocked neuropathic pain in mice.","method":"In silico docking screen, in vitro kinase assay, X-ray co-crystallography (chlortetracycline-EphB1), in vivo phosphorylation assay, neuropathic pain behavioral model","journal":"Proceedings of the National Academy of Sciences of the United States of America","confidence":"High","confidence_rationale":"Tier 1 / Strong — crystal structure + in vitro kinase assay + in vivo validation, multiple orthogonal methods","pmids":["33627480"],"is_preprint":false},{"year":2014,"finding":"EphB1 expressed in the striatal anlage signals to ephrin-B3 on migrating cortical interneurons and striatal neurons via reverse signaling with opposite effects: in cortical interneurons, EphB1-ephrin-B3 reverse signaling phosphorylates Src and FAK causing repulsion; in striatal neurons expressing ephrin-B3, EphB1 reduces pSrc and pFAK causing cessation of migration. In ephrin-B3 knockout mice, cortical interneurons are misrouted into the striatum and striatal neurons over-migrate.","method":"In vitro migration assays, ephrin-B3 knockout mouse, immunofluorescence for pSrc/pFAK, in vivo cell tracking","journal":"Frontiers in cellular neuroscience","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — in vitro assay + knockout mouse + biochemical phosphorylation in single lab","pmids":["25100946"],"is_preprint":false},{"year":2015,"finding":"EphB1 intracellular signaling domains are required for corpus callosum and anterior commissure formation. Truncated EphB1/EphB2 mice lacking intracellular domains show partial/complete agenesis of corpus callosum and highly penetrant anterior commissure misprojection, indicating forward (and potentially reverse) intracellular signaling by EphB1/EphB2 is required for forebrain axon guidance.","method":"Knock-in truncated EphB1/EphB2 mice, axon tract tracing, immunohistochemistry","journal":"Developmental neurobiology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — genetic knock-in with defined axon guidance phenotype, single lab","pmids":["26148571"],"is_preprint":false},{"year":2015,"finding":"EphB1 ligand-dependent signaling activates the DNA damage response (DDR) cascade via p53 DNA binding, leading to activation of ATR, Chk1, p53, p21, p38, CDK1(Tyr15) and Bax, and downregulation of HSP27 and Bcl2. In EphB1-methylated AML cells, re-introduction of EphB1 enhanced the DDR cascade and enforced programmed cell death, identifying EphB1 as a tumor suppressor in AML.","method":"EphB1 re-expression in methylated AML cells, signaling pathway analysis by western blot, promoter methylation analysis, p53 DNA binding assay","journal":"Molecular cancer research : MCR","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — re-expression rescue + signaling cascade analysis, single lab","pmids":["25944917"],"is_preprint":false},{"year":2023,"finding":"Neddylation of EphB1 enhances its kinase activity by preventing its degradation, thereby promoting proliferation, migration, and activation of hepatic stellate cells. EphB1 expression and neddylation are both increased in activated hepatic stellate cells.","method":"Neddylation assay (immunoprecipitation), kinase activity assay, hepatic stellate cell functional assays (proliferation, migration)","journal":"International journal of molecular sciences","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — biochemical neddylation assay + kinase activity + functional cell assays, single lab","pmids":["36834826"],"is_preprint":false},{"year":2023,"finding":"CRC-associated somatic mutations in EphB1 reduce kinase activity and protein stability. CRC mutant EphB1 receptors inhibit STAT3 and ERK1/2 signaling (contrasting wild-type) and are unable to suppress migration of CRC cells or compartmentalize cells in ephrin-B1 co-culture assays, establishing a kinase-dependent tumor suppressor function.","method":"Purified recombinant WT and mutant EphB1 kinase domain assays, cell migration assay, cell compartmentalization assay, mammalian cell expression","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 1 / Moderate — in vitro kinase reconstitution with purified proteins + cell-based functional assays, multiple mutants","pmids":["37527777"],"is_preprint":false},{"year":2018,"finding":"SUMOylation of EphB1 at lysine residue 785 suppresses cell proliferation, anchorage-independent growth, and xenograft tumor growth. SUMOylated EphB1 represses activation of its downstream effector PKCγ. A reciprocal regulatory loop between PKCγ and EphB1 SUMOylation was identified.","method":"Ni2+-NTA pull-down and immunoprecipitation for SUMOylation, K785R mutant expression, soft agar colony formation, xenograft mouse model, PKCγ activation assay","journal":"Cellular physiology and biochemistry","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — biochemical SUMOylation assay + mutagenesis + in vivo tumor model, single lab","pmids":["29550816"],"is_preprint":false},{"year":2024,"finding":"Tumor cell-expressed EPHB1 interacts with platelet-expressed EFNB1 (ephrin-B1) in the liver metastatic niche of pancreatic ductal adenocarcinoma. This contact-dependent EPHB1-EFNB1 interaction activates platelets via reverse signaling through AKT signaling; activated platelets then release serotonin (5-HT) which promotes tumor growth.","method":"Gain-of-function and loss-of-function of Ephb1, tumor cell-platelet adhesion assay, recombinant protein treatments, Tph1-knockout mice (serotonin-deficient), mCherry niche-labeling system","journal":"Cancer communications (London, England)","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — genetic LOF/GOF + knockout mice + mechanistic pathway in single lab","pmids":["39648610"],"is_preprint":false},{"year":2024,"finding":"EphB1 in GABAergic neurons (not cortical excitatory neurons or endothelial cells) is required for proper long-range cortical glutamatergic axon guidance. Conditional EphB1 knockout in GABAergic cells (Vgat-Cre) reproduces the cortical axon guidance defects of global EphB1 KO, with misguided axon bundles containing co-mingled GABAergic and glutamatergic axons near blood vessels.","method":"Cell-type-specific conditional knockout (Vgat-Cre, Emx1-Cre, Tie2-Cre), axon tract tracing, immunohistochemistry","journal":"Development (Cambridge, England)","confidence":"High","confidence_rationale":"Tier 2 / Strong — cell-type-specific conditional KO with multiple Cre lines to dissect cell-autonomous requirement, in vivo axon tracing","pmids":["38345254"],"is_preprint":false},{"year":2025,"finding":"EphB1 in glutamatergic neurons of the ventral posteromedial thalamic nucleus (VPM) promotes emergence from anesthesia by activating the VPMGlu→primary somatosensory cortex pathway. EphrinB-EphB1 signaling excites VPMGlu neurons through NR2B phosphorylation at Tyr-1472, and disinhibits VPMGlu neurons through ubiquitin-mediated degradation of KCC2; these are identified as two independent mechanisms.","method":"Conditional knockout, in vivo electrophysiology, NR2B phosphorylation assays, KCC2 ubiquitination assay, chemogenetics/optogenetics to probe neural circuit","journal":"Science advances","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — in vivo KO + circuit-level electrophysiology + molecular mechanism, single lab","pmids":["41348875"],"is_preprint":false},{"year":2020,"finding":"TGF-β-activated Smad2 transcriptionally upregulates endogenous EphB1 expression in lung cancer cells. Ligand-independent (unphosphorylated) EphB1 promotes epithelial-mesenchymal transition (EMT) by upregulating CDH2 (N-cadherin) and increases migration and invasion, while ligand-dependent (phosphorylated) EphB1 (activated by ephrin-B2) inhibits migration and invasion.","method":"EphB1 phosphorylation mutant expression, western blot for Smad2 and CDH2, transwell migration/invasion assay, siRNA knockdown","journal":"Journal of Cancer","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — phosphorylation mutant, siRNA, and signaling analysis, single lab","pmids":["32368295"],"is_preprint":false},{"year":2025,"finding":"EphB1 promotes CREB phosphorylation via PI3K/AKT signaling in a cell-autonomous manner in hypothalamic neurons. EphB1 deficiency reduces hypothalamic CRH and TRH expression, leading to impaired thermogenesis and locomotion (but not food intake), and results in obesity. Intraventricular administration of TRH or CRH suppressed obesity in Ephb1 mutant mice.","method":"Forward genetic screen, hypothalamic tissue and primary cell signaling analysis, PI3K/AKT pathway inhibitors, intracerebroventricular peptide delivery, metabolic phenotyping","journal":"Obesity (Silver Spring, Md.)","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — genetic mutant + cell-autonomous signaling + pharmacological rescue, single lab","pmids":["40207393"],"is_preprint":false},{"year":2023,"finding":"Recurring EPHB1 mutations in colorectal and other cancers alter receptor signaling and cell compartmentalization. Ligand-binding domain mutations (C61Y, R90C, R170W), fibronectin domain mutation (R351L), and kinase domain mutation (D762N) reduce compartmentalization and ligand-induced receptor phosphorylation. Kinase domain mutations R743W and G821R enhance compartmentalization without altered phosphorylation. Phosphoproteome analysis linked reduced-compartmentalization mutants to PI3K pathway/PIK3C2B phosphorylation.","method":"Confocal microscopy compartmentalization assay, phospho-proteome analysis, stable expression of 15 mutants in CRC cells","journal":"Cell communication and signaling : CCS","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — systematic functional mutagenesis panel + phosphoproteomics, single lab","pmids":["38102712"],"is_preprint":false}],"current_model":"EphB1 is a receptor tyrosine kinase that, upon ligand (ephrin-B) binding, undergoes autophosphorylation and recruits a set of adaptor/effector proteins (Nck via pTyr594, Grb7 via pTyr928, p52Shc/c-Src via Tyr600/778, Grb2, Cbl, PTEN/NHERF1, Porf-2/RhoGAP, Caskin1) to activate downstream JNK, ERK/MAPK, and Rac1-GTPase pathways that regulate cell migration, adhesion, and actin cytoskeletal dynamics; it controls neural circuit formation (ipsilateral retinal projection via ephrin-B2 repulsion at the optic chiasm, dopaminergic pathway specification, cortical axon guidance through GABAergic neurons), modulates synaptic plasticity and pain processing via NR2B/NMDA receptor phosphorylation, regulates caveolae biogenesis through constitutive Cav-1 interaction, and is post-translationally regulated by Cbl-mediated ubiquitination/lysosomal degradation, SUMOylation at K785 (suppressing PKCγ), and neddylation (enhancing kinase activity), with its tumor-suppressive kinase function frequently impaired by somatic mutations in colorectal and other cancers."},"narrative":{"mechanistic_narrative":"EphB1 is an ephrin-B-activated receptor tyrosine kinase that, upon ligand engagement and autophosphorylation, assembles phosphotyrosine-dependent adaptor complexes to control cell adhesion, migration, and actin cytoskeletal dynamics [PMID:10205170, PMID:9430661, PMID:12925710]. Activated EphB1 recruits the SH2-adaptor Nck through phospho-Tyr594 to drive JNK activation and cell attachment, and the Nck–paxillin/FAK axis with c-Src-dependent paxillin phosphorylation to promote migration [PMID:9430661, PMID:15107421]; it also engages Grb2/p52Shc and Grb7 through distinct phosphotyrosines (Tyr600/Tyr778, Tyr928) to activate ERK/MAPK and chemotaxis [PMID:12925710, PMID:12223469], and recruits the scaffold Caskin1 via Nck [PMID:23181695]. Forward signaling brakes axon outgrowth by binding the RhoGAP Porf-2 to deactivate Rac1 [PMID:29938386], while PTEN, delivered through NHERF1, antagonizes EphB1-dependent adhesion and chemotaxis [PMID:23118026]. EphB1 is a principal determinant of neural circuit wiring: it functions as the preferred receptor for ephrin-B2 at the optic chiasm, where its kinase-competent forward signaling repels ventrotemporal retinal ganglion cell axons to establish the ipsilateral projection downstream of the Zic2 transcription factor [PMID:12971893, PMID:18524895, PMID:19295152, PMID:22103419], and it directs neural crest migration, dopaminergic pathway specification, and forebrain commissural and long-range cortical axon guidance—the latter cell-autonomously through GABAergic neurons [PMID:9259557, PMID:10066262, PMID:17561836, PMID:26148571, PMID:38345254]. EphB1 localizes to caveolae and is required for caveolae biogenesis by constitutively binding and stabilizing caveolin-1 against ubiquitin-mediated degradation [PMID:16723736, PMID:32238105]. Its abundance and activity are tuned post-translationally—by Cbl-mediated ubiquitination and lysosomal degradation [PMID:18034775], SUMOylation at Lys785 that suppresses PKCγ and tumor growth [PMID:29550816], and neddylation that stabilizes the receptor and enhances kinase activity [PMID:36834826]. Kinase-dependent EphB1 acts as a tumor suppressor: cancer-associated somatic mutations reduce its activity, stability, and ability to compartmentalize cells, and re-expression of EphB1 in silenced leukemia cells enforces a p53-driven DNA-damage/apoptotic program [PMID:25944917, PMID:37527777, PMID:38102712].","teleology":[{"year":1998,"claim":"Established the first proximal EphB1 effector linkage by showing how a defined receptor phosphotyrosine couples kinase activation to a downstream MAP kinase and an adhesive cell response.","evidence":"Yeast two-hybrid, Co-IP, Y594F mutagenesis, JNK and cell attachment assays","pmids":["9430661"],"confidence":"High","gaps":["Did not define the kinase chain from Nck to JNK","Physiological cell context of the attachment response not addressed"]},{"year":1999,"claim":"Defined EphB1 as a ligand-density sensor whose kinase signaling, not mechanical tethering, converts ephrin-B engagement into integrin-mediated attachment.","evidence":"Reconstituted ephrin-B1 surface-density assay with integrin blocking and signaling-defective EphB1 mutants","pmids":["10205170"],"confidence":"High","gaps":["Mechanism coupling EphB1 to specific integrins not resolved","Tested in endothelial and transfected HEK cells only"]},{"year":2002,"claim":"Mapped a second adaptor branch by identifying Grb7 as an autophosphorylation-dependent EphB1 partner and substrate that drives motility.","evidence":"Yeast two-hybrid, Co-IP, Tyr928 mutagenesis, kinase and migration assays","pmids":["12223469"],"confidence":"High","gaps":["Downstream effectors of Grb7 in EphB1 migration unresolved"]},{"year":2003,"claim":"Resolved the EphB1 migration pathway by linking Tyr600/Tyr778, c-Src, p52Shc and ERK to chemotaxis as distinct from adhesion.","evidence":"Co-IP, Y600F/Y778F mutagenesis, dominant-negative Src, PD98059/PP2 inhibitors, migration vs adhesion assays","pmids":["12925710"],"confidence":"High","gaps":["Spatial coordination of parallel adhesion and migration outputs not defined"]},{"year":2003,"claim":"Demonstrated that EphB1 forward signaling drives a specific developmental axon-sorting decision, establishing its role in binocular circuit formation.","evidence":"EphB1 knockout mice, expression mapping, ephrin-B2 function-blocking in semiintact visual system","pmids":["12971893"],"confidence":"High","gaps":["Intracellular effectors mediating chiasm repulsion not identified in this study"]},{"year":2004,"claim":"Built the EphB1→Nck→paxillin/FAK→Src migration complex, connecting the juxtamembrane adaptor to focal-adhesion substrate phosphorylation.","evidence":"Co-IP, paxillin Y31F/Y118F and EphB1 Y594F mutagenesis, dominant-negative Src, PP2, migration assay","pmids":["15107421"],"confidence":"High","gaps":["Whether the complex acts at nascent versus mature adhesions unresolved"]},{"year":2006,"claim":"Placed EphB1 in caveolae and tied its membrane targeting and ERK output to a caveolin-binding motif in the kinase domain.","evidence":"Fractionation, Co-IP, Cav-1 scaffolding-domain mutant, confocal microscopy, ERK assays","pmids":["16723736"],"confidence":"Medium","gaps":["Did not determine whether caveolae are required for all EphB1 signaling outputs"]},{"year":2007,"claim":"Defined the negative-feedback arm of EphB1 signaling through Cbl-mediated ubiquitination and lysosomal turnover.","evidence":"Co-IP, GST pull-down, Cbl ligase-dead and EphB1-K652R mutants, ubiquitination assay, bafilomycin, PP2","pmids":["18034775"],"confidence":"High","gaps":["Kinetics of receptor recycling versus degradation not quantified"]},{"year":2008,"claim":"Identified the transcriptional control point upstream of EphB1 by showing Zic2 induces functional EphB1 to convert axon behavior to ephrin-B2 avoidance.","evidence":"Retinal explant electroporation, avoidance assays on ephrin-B2, growth-cone immunofluorescence","pmids":["18524895"],"confidence":"High","gaps":["Direct versus indirect regulation of the EphB1 promoter by Zic2 not established"]},{"year":2009,"claim":"Showed EphB1 is both necessary and sufficient for ipsilateral redirection and that its specificity resides in extracellular and juxtamembrane domains.","evidence":"In utero retinal electroporation, EphB1-EphB2 chimeric receptors, in vivo tracing","pmids":["19295152"],"confidence":"High","gaps":["Molecular basis of domain-specific specificity over EphB2 not defined at residue level"]},{"year":2011,"claim":"Distinguished forward from reverse signaling, proving EphB1 intracellular signaling drives the ipsilateral projection.","evidence":"Intracellular-truncated EphB1 knock-in mice, axon tracing, EphB/ephrin-B genetic combinations","pmids":["22103419"],"confidence":"High","gaps":["Effector cascade downstream of forward signal in RGCs not fully mapped"]},{"year":2012,"claim":"Extended EphB1 adaptor wiring by identifying Caskin1 as a Nck-bridged substrate undergoing phosphorylation-induced conformational change.","evidence":"Co-IP, mass-spectrometry phosphosite mapping, CD spectroscopy","pmids":["23181695"],"confidence":"Medium","gaps":["Functional consequence of Caskin1 phosphorylation for EphB1 output untested"]},{"year":2012,"claim":"Revealed PTEN/NHERF1 as a phosphatase brake that disrupts the PTEN-Cbl complex and limits EphB1-driven adhesion and chemotaxis.","evidence":"Co-IP, PTEN and NHERF1 siRNA, PTEN phosphatase mutants, adhesion and chemotaxis assays","pmids":["23118026"],"confidence":"Medium","gaps":["In vivo relevance of the PTEN-Cbl switch not addressed"]},{"year":2018,"claim":"Identified the RhoGAP Porf-2 as the link between EphB1 forward signaling and Rac1 inactivation in axon-growth braking.","evidence":"Co-IP, GTP-Rac1 pull-down, GAP-domain mutagenesis, axon growth measurement","pmids":["29938386"],"confidence":"Medium","gaps":["Recruitment mechanism of Porf-2 to activated EphB1 unresolved"]},{"year":2018,"claim":"Established SUMOylation at Lys785 as a tumor-suppressive post-translational switch that represses PKCγ.","evidence":"Ni-NTA/IP SUMOylation assay, K785R mutant, soft-agar and xenograft assays, PKCγ activation assay","pmids":["29550816"],"confidence":"Medium","gaps":["SUMO ligase responsible not identified","Relationship to kinase activity not defined"]},{"year":2020,"claim":"Demonstrated that EphB1 constitutively stabilizes caveolin-1 and is required for caveolae biogenesis in endothelial cells.","evidence":"Super-resolution microscopy, FRET, Co-IP, CSD-motif deletion mutant, EphB1-KO ECs, EM caveolae counting","pmids":["32238105"],"confidence":"High","gaps":["Physiological vascular consequences of caveolae loss not assessed"]},{"year":2020,"claim":"Uncovered phosphorylation-state-dependent dual function: ligand-independent EphB1 promotes EMT while activated EphB1 inhibits invasion, downstream of TGF-β/Smad2.","evidence":"EphB1 phosphorylation mutants, Smad2/CDH2 western blot, transwell assays, siRNA","pmids":["32368295"],"confidence":"Medium","gaps":["Mechanism by which unphosphorylated EphB1 upregulates N-cadherin unclear"]},{"year":2021,"claim":"Provided a structural and pharmacological handle by co-crystallizing a tetracycline in the EphB1 ATP pocket and blocking neuropathic pain in vivo.","evidence":"In silico docking, in vitro kinase assay, X-ray co-crystallography, in vivo phosphorylation and pain model","pmids":["33627480"],"confidence":"High","gaps":["Selectivity over related Eph kinases not fully characterized"]},{"year":2023,"claim":"Established EphB1 kinase activity as essential for its tumor-suppressor function, with cancer mutations crippling activity, stability and cell compartmentalization.","evidence":"Purified WT/mutant kinase-domain assays, migration and compartmentalization assays in CRC cells","pmids":["37527777"],"confidence":"High","gaps":["In vivo tumor-suppressor validation of specific mutants not shown"]},{"year":2023,"claim":"Showed neddylation stabilizes EphB1 and enhances kinase activity to promote hepatic stellate cell activation.","evidence":"Neddylation IP, kinase activity assay, proliferation and migration assays","pmids":["36834826"],"confidence":"Medium","gaps":["Neddylation site and responsible ligase not mapped"]},{"year":2023,"claim":"Systematically parsed cancer-mutation effects, linking reduced-compartmentalization mutants to altered PI3K-pathway phosphorylation.","evidence":"Confocal compartmentalization assay and phosphoproteomics across 15 mutants in CRC cells","pmids":["38102712"],"confidence":"Medium","gaps":["Causal role of PIK3C2B phosphorylation in compartmentalization not tested"]},{"year":2024,"claim":"Pinpointed the cell-autonomous requirement for EphB1 in GABAergic neurons for long-range cortical glutamatergic axon guidance.","evidence":"Cell-type-specific conditional knockout (Vgat-Cre, Emx1-Cre, Tie2-Cre), axon tracing, IHC","pmids":["38345254"],"confidence":"High","gaps":["Molecular signal between GABAergic EphB1 and glutamatergic axons not defined"]},{"year":2024,"claim":"Revealed a tumor-platelet axis in which tumor EPHB1 activates platelets via ephrin-B1 reverse signaling to release pro-tumor serotonin.","evidence":"Ephb1 GOF/LOF, tumor-platelet adhesion assay, recombinant proteins, Tph1-knockout mice, niche labeling","pmids":["39648610"],"confidence":"Medium","gaps":["Generalizability beyond pancreatic liver-metastatic niche untested"]},{"year":2025,"claim":"Defined dual molecular mechanisms—NR2B Tyr1472 phosphorylation and KCC2 degradation—by which EphB1 drives thalamocortical excitation and anesthesia emergence.","evidence":"Conditional KO, in vivo electrophysiology, NR2B phosphorylation and KCC2 ubiquitination assays, chemo/optogenetics","pmids":["41348875"],"confidence":"Medium","gaps":["Relative contribution of the two mechanisms in vivo not quantified"]},{"year":2025,"claim":"Linked EphB1 to hypothalamic energy homeostasis via cell-autonomous PI3K/AKT/CREB control of CRH/TRH and thermogenesis.","evidence":"Forward genetic screen, PI3K/AKT inhibitors, ICV peptide rescue, metabolic phenotyping","pmids":["40207393"],"confidence":"Medium","gaps":["Whether the phenotype requires ligand engagement or kinase activity unresolved"]},{"year":null,"claim":"How EphB1's many context-dependent outputs—forward vs reverse signaling, kinase-dependent tumor suppression vs ligand-independent EMT promotion, and tissue-specific effector selection—are integrated at a structural and regulatory level remains unresolved.","evidence":"","pmids":[],"confidence":"Medium","gaps":["No unified model linking post-translational modifications (Cbl, SUMO, neddylation) to effector choice","Determinants of receptor specificity over other EphB receptors in vivo undefined","Structural basis of compartmentalization versus signaling outputs unknown"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0140096","term_label":"catalytic activity, acting on a protein","supporting_discovery_ids":[1,2,3,5,25]},{"term_id":"GO:0016740","term_label":"transferase activity","supporting_discovery_ids":[3,20,25]},{"term_id":"GO:0060089","term_label":"molecular transducer activity","supporting_discovery_ids":[0,8,13]},{"term_id":"GO:0060090","term_label":"molecular adaptor activity","supporting_discovery_ids":[1,15]}],"localization":[{"term_id":"GO:0005886","term_label":"plasma membrane","supporting_discovery_ids":[6,19]},{"term_id":"GO:0031410","term_label":"cytoplasmic vesicle","supporting_discovery_ids":[6,19]}],"pathway":[{"term_id":"R-HSA-162582","term_label":"Signal Transduction","supporting_discovery_ids":[1,2,5]},{"term_id":"R-HSA-1266738","term_label":"Developmental Biology","supporting_discovery_ids":[8,12,13,28]},{"term_id":"R-HSA-1643685","term_label":"Disease","supporting_discovery_ids":[23,25,32]},{"term_id":"R-HSA-112316","term_label":"Neuronal 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The signaling pathway downstream of the receptor is referred to as forward signaling while the signaling pathway downstream of the ephrin ligand is referred to as reverse signaling. Cognate/functional ephrin ligands for this receptor include EFNB1, EFNB2 and EFNB3. During nervous system development, regulates retinal axon guidance redirecting ipsilaterally ventrotemporal retinal ganglion cells axons at the optic chiasm midline. This probably requires repulsive interaction with EFNB2. In the adult nervous system together with EFNB3, regulates chemotaxis, proliferation and polarity of the hippocampus neural progenitors. In addition to its role in axon guidance also plays an important redundant role with other ephrin-B receptors in development and maturation of dendritic spines and synapse formation. May also regulate angiogenesis. More generally, may play a role in targeted cell migration and adhesion. Upon activation by EFNB1 and probably other ephrin-B ligands activates the MAPK/ERK and the JNK signaling cascades to regulate cell migration and adhesion respectively. Involved in the maintenance of the pool of satellite cells (muscle stem cells) by promoting their self-renewal and reducing their activation and differentiation (By similarity)","subcellular_location":"Cell membrane; Early endosome membrane; Cell projection, dendrite","url":"https://www.uniprot.org/uniprotkb/P54762/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/EPHB1","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":[],"url":"https://opencell.sf.czbiohub.org/search/EPHB1","total_profiled":1310},"omim":[{"mim_id":"609481","title":"ISL2 TRANSCRIPTION FACTOR, LIM/HOMEODOMAIN; ISL2","url":"https://www.omim.org/entry/609481"},{"mim_id":"601581","title":"NEURONAL CELL ADHESION MOLECULE; NRCAM","url":"https://www.omim.org/entry/601581"},{"mim_id":"601381","title":"EPHRIN A3; EFNA3","url":"https://www.omim.org/entry/601381"},{"mim_id":"601380","title":"EPHRIN A4; EFNA4","url":"https://www.omim.org/entry/601380"},{"mim_id":"600600","title":"EPHRIN RECEPTOR EphB1; EPHB1","url":"https://www.omim.org/entry/600600"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"Supported","locations":[{"location":"Endoplasmic reticulum","reliability":"Supported"},{"location":"Plasma membrane","reliability":"Supported"},{"location":"Cytosol","reliability":"Additional"}],"tissue_specificity":"Tissue enhanced","tissue_distribution":"Detected in many","driving_tissues":[{"tissue":"brain","ntpm":19.3}],"url":"https://www.proteinatlas.org/search/EPHB1"},"hgnc":{"alias_symbol":["Hek6"],"prev_symbol":["EPHT2"]},"alphafold":{"accession":"P54762","domains":[{"cath_id":"2.60.120.260","chopping":"19-197","consensus_level":"high","plddt":84.9964,"start":19,"end":197},{"cath_id":"2.60.40.1770","chopping":"200-256","consensus_level":"medium","plddt":88.2319,"start":200,"end":256},{"cath_id":"2.60.40.10","chopping":"323-429","consensus_level":"medium","plddt":92.7064,"start":323,"end":429},{"cath_id":"2.60.40.10","chopping":"442-532","consensus_level":"high","plddt":88.2515,"start":442,"end":532},{"cath_id":"3.30.200.20","chopping":"598-699","consensus_level":"high","plddt":86.9715,"start":598,"end":699},{"cath_id":"1.10.510.10","chopping":"704-889","consensus_level":"high","plddt":87.3124,"start":704,"end":889},{"cath_id":"1.10.150.50","chopping":"911-979","consensus_level":"high","plddt":81.3207,"start":911,"end":979}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/P54762","model_url":"https://alphafold.ebi.ac.uk/files/AF-P54762-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-P54762-F1-predicted_aligned_error_v6.png","plddt_mean":83.62},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=EPHB1","jax_strain_url":"https://www.jax.org/strain/search?query=EPHB1"},"sequence":{"accession":"P54762","fasta_url":"https://rest.uniprot.org/uniprotkb/P54762.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/P54762/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/P54762"}},"corpus_meta":[{"pmid":"9259557","id":"PMC_9259557","title":"The 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pathology","url":"https://pubmed.ncbi.nlm.nih.gov/25120806","citation_count":5,"is_preprint":false},{"pmid":"23118026","id":"PMC_23118026","title":"Phosphatase and tensin homolog regulates stability and activity of EphB1 receptor.","date":"2012","source":"FASEB journal : official publication of the Federation of American Societies for Experimental Biology","url":"https://pubmed.ncbi.nlm.nih.gov/23118026","citation_count":5,"is_preprint":false},{"pmid":"24606480","id":"PMC_24606480","title":"EphB1 and Ephrin-B, new potential biomarkers for squamous cell/adenosquamous carcinomas and adenocarcinomas of the gallbladder.","date":"2014","source":"Asian Pacific journal of cancer prevention : APJCP","url":"https://pubmed.ncbi.nlm.nih.gov/24606480","citation_count":4,"is_preprint":false},{"pmid":"39648610","id":"PMC_39648610","title":"Reciprocal tumor-platelet interaction through the EPHB1-EFNB1 axis in the liver metastatic niche promotes metastatic tumor outgrowth in pancreatic ductal adenocarcinoma.","date":"2024","source":"Cancer communications (London, England)","url":"https://pubmed.ncbi.nlm.nih.gov/39648610","citation_count":4,"is_preprint":false},{"pmid":"38345254","id":"PMC_38345254","title":"EphB1 controls long-range cortical axon guidance through a cell non-autonomous role in GABAergic cells.","date":"2024","source":"Development (Cambridge, England)","url":"https://pubmed.ncbi.nlm.nih.gov/38345254","citation_count":4,"is_preprint":false},{"pmid":"21763378","id":"PMC_21763378","title":"Associations of EPHB1 polymorphisms with hepatocellular carcinoma in the Korean population.","date":"2011","source":"Human immunology","url":"https://pubmed.ncbi.nlm.nih.gov/21763378","citation_count":4,"is_preprint":false},{"pmid":"38102712","id":"PMC_38102712","title":"Recurring EPHB1 mutations in human cancers alter receptor signalling and compartmentalisation of colorectal cancer cells.","date":"2023","source":"Cell communication and signaling : 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Activation-competent but signaling-defective EphB1 point mutants failed to stimulate ephrin-B1-dependent attachment, demonstrating requirement for EphB1 kinase signaling.\",\n      \"method\": \"Reconstituted ephrin-B1 surface density assay, integrin-blocking antibodies, EphB1 point mutants in transfection experiments, cell attachment assays\",\n      \"journal\": \"The EMBO journal\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — in vitro reconstitution with point-mutant validation, single lab but multiple orthogonal methods (density assay + integrin blocking + mutagenesis)\",\n      \"pmids\": [\"10205170\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 1998,\n      \"finding\": \"The adapter protein Nck binds directly to the juxtamembrane region of EphB1 via its SH2 domain, requiring phospho-Tyr594. Ligand (ephrin-B1/Fc) activation of EphB1 recruits Nck to native receptor complexes and activates c-Jun kinase (JNK/SAPK). Mutant EphB1-Y594F blocks Nck recruitment, attenuates JNK activation, and blocks cell attachment responses.\",\n      \"method\": \"Yeast two-hybrid cloning, Co-immunoprecipitation, site-directed mutagenesis (Y594F), JNK kinase assay, cell attachment assay\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1-2 / Moderate — yeast two-hybrid + Co-IP + mutagenesis + kinase assay in single lab with multiple orthogonal methods\",\n      \"pmids\": [\"9430661\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2003,\n      \"finding\": \"Activated EphB1 recruits adaptor proteins Grb2 and p52Shc, promotes c-Src-dependent tyrosine phosphorylation of p52Shc, and activates MAPK/ERK to drive cell chemotaxis. EphB1 Tyr600 and Tyr778 are required for interaction with c-Src and p52Shc. Phosphorylation of p52Shc by c-Src is required for its recruitment to EphB1 complexes via its phosphotyrosine binding domain. ERK and Src inhibitors abolished EphB1-mediated migration but not adhesion.\",\n      \"method\": \"Co-immunoprecipitation, site-directed mutagenesis (Y600F, Y778F), dominant-negative c-Src expression, pharmacological inhibitors (PD98059, PP2), cell migration and adhesion assays\",\n      \"journal\": \"The Journal of cell biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1-2 / Moderate — reciprocal Co-IP, mutagenesis, dominant-negative, and pharmacological approaches in single lab\",\n      \"pmids\": [\"12925710\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2002,\n      \"finding\": \"Grb7 binds to the cytoplasmic domain of EphB1 via its SH2 domain in a manner requiring EphB1 autophosphorylation. Tyr-928 of EphB1 is the primary Grb7 binding site. EphB1 can phosphorylate Grb7, and mutations Y928F or Y594F reduced this activity. Co-expression of Grb7 with EphB1 enhanced cell motility, while the Grb7 SH2 domain alone abolished EphB1-stimulated migration.\",\n      \"method\": \"Yeast two-hybrid screening, Co-immunoprecipitation, site-directed mutagenesis, kinase assay, cell migration assay on extracellular matrix\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — yeast two-hybrid + Co-IP + mutagenesis + functional migration assay in single lab\",\n      \"pmids\": [\"12223469\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2001,\n      \"finding\": \"The kinase-inactive EphB6 receptor undergoes transphosphorylation upon ligand (ephrin-B1) stimulation when co-expressed with catalytically active EphB1. EphB1 and EphB6 form a stable hetero-complex; EphB1-induced EphB6 phosphorylation requires EphB1 catalytic activity. The proto-oncogene c-Cbl was identified as a constitutive EphB6-binding protein requiring a functional phosphotyrosine binding domain.\",\n      \"method\": \"Co-immunoprecipitation, overexpression of kinase-dead EphB1 mutants, ligand stimulation assays\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — reciprocal Co-IP with mutant controls, single lab, two methods\",\n      \"pmids\": [\"11713248\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2004,\n      \"finding\": \"Activated EphB1 induces c-Src-dependent tyrosine phosphorylation of paxillin at Tyr-31 and Tyr-118, and is recruited to paxillin-FAK complexes. Cells expressing paxillin Y31F/Y118F or paxillin ΔLD4 show reduced EphB1-dependent migration. Nck binding site mutation (Y594F) disrupts the Nck-paxillin-EphB1 complex, establishing a signaling complex: EphB1 → Nck → paxillin/FAK → c-Src-mediated paxillin phosphorylation → cell migration.\",\n      \"method\": \"Co-immunoprecipitation, site-directed mutagenesis (paxillin Y31F/Y118F, EphB1 Y594F), dominant-negative c-Src, Src inhibitor PP2, cell migration assay\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1-2 / Moderate — mutagenesis of both receptor and substrate, Co-IP, dominant-negative, inhibitor validation in single lab\",\n      \"pmids\": [\"15107421\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2006,\n      \"finding\": \"EphB1 receptors localize to caveolae and directly interact with caveolin-1 (Cav-1) upon ligand stimulation. This interaction requires the caveolin-binding motif within the EphB1 kinase domain, which is also required for correct membrane targeting of EphB1. Overexpression of a Cav-1 mutant lacking the scaffolding domain abolishes EphB1-Cav-1 interaction and EphB1-mediated ERK activation.\",\n      \"method\": \"Subcellular fractionation, co-immunoprecipitation, caveolin-1 scaffolding domain mutant overexpression, confocal microscopy, ERK activation assays\",\n      \"journal\": \"Journal of cell science\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — Co-IP, fractionation, dominant-negative mutant, and signaling assay in single lab\",\n      \"pmids\": [\"16723736\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2007,\n      \"finding\": \"Ligand (ephrin-B1) stimulation of EphB1 leads to Cbl recruitment to EphB1 via Cbl's tyrosine kinase-binding domain, followed by Src-dependent Cbl phosphorylation, EphB1 ubiquitination, and lysosomal degradation of EphB1. Overexpression of wild-type but not the ligase-dead 70Z Cbl mutant enhanced EphB1 ubiquitination and degradation. Kinase-dead EphB1-K652R was resistant to Cbl-mediated degradation.\",\n      \"method\": \"Co-immunoprecipitation, GST pull-down, overexpression of Cbl mutants, ubiquitination assay, lysosomal inhibitor (bafilomycin) treatment, Src inhibitor (PP2)\",\n      \"journal\": \"Traffic (Copenhagen, Denmark)\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1-2 / Moderate — GST pull-down + Co-IP + ubiquitination assay + multiple mutants in single lab\",\n      \"pmids\": [\"18034775\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2003,\n      \"finding\": \"Ephrin-B2 is expressed at the optic chiasm midline and selectively repels ventrotemporal retinal ganglion cell (RGC) axons that express EphB1. EphB1 is found exclusively in retinal regions giving rise to the ipsilateral projection. EphB1 null mice exhibit a dramatically reduced ipsilateral retinal projection, and blocking ephrin-B2 function in vitro eliminates the inhibitory effect on chiasm cells and abolishes ipsilateral projection in semiintact preparation.\",\n      \"method\": \"Immunohistochemistry, EphB1 knockout mouse, in vitro axon growth assays with function-blocking reagents, semiintact visual system preparation\",\n      \"journal\": \"Neuron\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — genetic knockout + in vitro blocking assays + expression localization, replicated in subsequent studies\",\n      \"pmids\": [\"12971893\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 1997,\n      \"finding\": \"EphB1 (and EphA4) expressed in migrating third arch neural crest cells interact with ephrin-B2 in second arch neural crest/mesoderm to restrict intermingling and direct targeted migration. Inhibition of EphB1/EphA4 function using truncated receptors leads to abnormal third arch neural crest migration into second and fourth arch territories; ectopic ephrin-B2 overexpression scatters third arch cells.\",\n      \"method\": \"Dominant-negative truncated receptor expression in Xenopus embryos, ectopic ephrin-B2 overexpression, in situ hybridization, cell lineage tracing\",\n      \"journal\": \"Current biology : CB\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — dominant-negative and gain-of-function in Xenopus embryos, single lab\",\n      \"pmids\": [\"9259557\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 1999,\n      \"finding\": \"EphB1 and its ligand ephrin-B2 are expressed in complementary patterns in midbrain dopaminergic neurons and their targets. Ephrin-B2 selectively inhibits neurite outgrowth and induces cell loss of substantia nigra (but not ventral tegmental) dopaminergic neurons, suggesting EphB1/ephrin-B2 specifies distinct dopaminergic pathways. Ephrin-B2 expression is upregulated by cocaine and amphetamine in adults.\",\n      \"method\": \"Neurite outgrowth inhibition assay, cell survival assay, in situ hybridization, immunohistochemistry\",\n      \"journal\": \"The Journal of neuroscience\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — in vitro functional assays with specific ligand, expression pattern analysis, single lab\",\n      \"pmids\": [\"10066262\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2008,\n      \"finding\": \"Zic2 transcription factor regulates EphB1 expression in retinal ganglion cells. Ectopic delivery of Zic2 into non-VT retinal explants induces EphB1 mRNA and protein expression. The upregulated EphB1 localizes to growth cones and is functional, converting RGC axon behavior from extension onto to avoidance of ephrin-B2 substrates, demonstrating Zic2→EphB1 axis controls ipsilateral projection.\",\n      \"method\": \"Retinal explant electroporation, in vitro axon growth/avoidance assays on ephrin-B2 substrates, immunofluorescence for EphB1 protein in growth cones\",\n      \"journal\": \"The Journal of neuroscience\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — gain-of-function with functional readout, protein localization, multiple assays in single lab\",\n      \"pmids\": [\"18524895\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2009,\n      \"finding\": \"EphB1 is specifically required and sufficient to drive the ipsilateral retinal projection. In utero retinal electroporation of EphB1 redirects normally-crossed RGCs to ipsilateral trajectory. EphB2, despite high similarity, is far less efficient. EphB1-EphB2 chimeric receptor analysis reveals that both extracellular and juxtamembrane domains of EphB1 are specifically required for ipsilateral redirection.\",\n      \"method\": \"In utero retinal electroporation, chimeric receptor expression, in vivo axon tracing\",\n      \"journal\": \"The Journal of neuroscience\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — in vivo gain-of-function with chimeric receptor dissection, domain-specific analysis\",\n      \"pmids\": [\"19295152\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2011,\n      \"finding\": \"EphB1 forward signaling (into the EphB1-expressing cell) is specifically required for ipsilateral retinal axon projection; reverse signaling through the EphB1 extracellular domain is not required. Knock-in mice expressing intracellular-truncated EphB1 (EphB1T-lacZ) fail to form the ipsilateral projection, phenocopying the EphB1 null. EphB1 is the preferred receptor for ephrin-B2 (and less so ephrin-B1) at the optic chiasm.\",\n      \"method\": \"Knock-in mouse (intracellular truncation), retinal axon tracing, genetic analysis of EphB/ephrin-B mutant combinations\",\n      \"journal\": \"The European journal of neuroscience\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — clean knock-in distinguishing forward vs. reverse signaling, in vivo tracing\",\n      \"pmids\": [\"22103419\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2008,\n      \"finding\": \"EphB1 null mice show a 40% decrease in neuron number and volume in substantia nigra pars reticulata (SNr), but not in pars compacta TH+ neurons. EphB1 is expressed in SNr (not SNc) neurons. Loss of EphB1 results in spontaneous locomotor hyperactivity, linking EphB1-dependent SNr development to locomotor function.\",\n      \"method\": \"EphB1 knockout mice, stereological neuron counting, in situ hybridization, beta-galactosidase reporter, open-field behavioral testing\",\n      \"journal\": \"The European journal of neuroscience\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — genetic knockout with anatomical and behavioral readout, single lab\",\n      \"pmids\": [\"17561836\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2012,\n      \"finding\": \"EphB1 recruits scaffold protein Caskin1 via the adaptor Nck: EphB1 activation leads to Nck SH2 binding to phosphotyrosine on EphB1, while Nck SH3 domains interact with the proline-rich domain of Caskin1. This complex formation results in tyrosine phosphorylation of Caskin1 at Tyr296 and Tyr336 (identified by mass spectrometry), and phosphorylation causes structural changes in the Caskin1 SH3 domain detected by CD spectroscopy.\",\n      \"method\": \"Co-immunoprecipitation, mass spectrometry phosphosite identification, CD spectroscopy\",\n      \"journal\": \"Cell communication and signaling : CCS\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — Co-IP + MS phosphosite + structural CD spectroscopy in single lab\",\n      \"pmids\": [\"23181695\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2012,\n      \"finding\": \"PTEN is constitutively associated with c-Cbl, protecting Cbl from degradation. EphB1 stimulation triggers PTEN dephosphorylation (~50% on Ser/Thr) and disruption of the PTEN-Cbl complex, requiring PTEN protein phosphatase activity. Both PTEN and Cbl independently translocate to EphB1, with PTEN associating via scaffold protein NHERF1. PTEN lipid phosphatase activity impairs EphB1-dependent cell adhesion and chemotaxis.\",\n      \"method\": \"Co-immunoprecipitation, PTEN and NHERF1 siRNA knockdown, PTEN phosphatase mutants, cell adhesion and chemotaxis assays\",\n      \"journal\": \"FASEB journal\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — Co-IP, siRNA, and phosphatase-dead mutants with functional assays, single lab\",\n      \"pmids\": [\"23118026\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"EphB1 is upregulated in injured motor neurons and can activate astrocytes through ephrin-B1-mediated stimulation of STAT3. EphB1-induced astrocyte activation produces a protective and anti-inflammatory transcriptional signature distinct from IL-6-induced responses. This EphB1-ephrin-B1-STAT3 pathway is disrupted in human stem cell-derived astrocytes and mouse models of ALS.\",\n      \"method\": \"Transcriptional analysis of EphB1-stimulated astrocytes, STAT3 pathway analysis, human iPSC-derived astrocytes, ALS mouse models\",\n      \"journal\": \"Nature communications\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — multiple model systems (human iPSC and mouse), pathway analysis, single lab\",\n      \"pmids\": [\"29079839\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"The RhoGAP protein Porf-2 is an intracellular mediator of EphB1 signaling in axon growth inhibition. EphB1 binds and regulates Porf-2 upon ephrin-B stimulation. The activated EphB1 forward signal deactivates Rac1 through the GAP domain of Porf-2, which inhibits growth cone expansion and brakes axon outgrowth.\",\n      \"method\": \"Co-immunoprecipitation of EphB1-Porf-2, Rac1 activity assays (GTP-Rac1 pull-down), axon growth measurement, GAP domain mutagenesis\",\n      \"journal\": \"Cellular and molecular life sciences : CMLS\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — Co-IP, Rac1 activity assay, functional axon growth readout in single lab\",\n      \"pmids\": [\"29938386\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"EphB1 constitutively interacts with caveolin-1 (Cav-1) in endothelial cells. Ephrin-B1 activation of EphB1 induces EphB1 phosphorylation, uncouples EphB1 from Cav-1, and promotes Src-dependent Cav-1 phosphorylation. Deletion of the CSD-binding motif in EphB1 prevents Cav-1 interaction and Cav-1 phosphorylation. EphB1-/- ECs show markedly reduced Cav-1 expression and caveolae numbers due to Cav-1 ubiquitination and degradation when not bound to EphB1.\",\n      \"method\": \"Super-resolution microscopy, FRET, co-immunoprecipitation, EphB1 CSD-motif deletion mutant, EphB1 knockout endothelial cells, electron microscopy caveolae counting\",\n      \"journal\": \"Molecular biology of the cell\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1-2 / Strong — FRET + Co-IP + mutagenesis + knockout ECs + structural imaging, multiple orthogonal methods\",\n      \"pmids\": [\"32238105\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"Tetracycline antibiotics demeclocycline, chlortetracycline, and minocycline inhibit EphB1 kinase activity at low micromolar concentrations by binding to the ATP-binding catalytic domain. Co-crystallization of chlortetracycline with EphB1 confirmed binding to the ATP-binding domain. In vivo, the three-tetracycline combination inhibited EphB1 phosphorylation in brain, spinal cord, and DRG and blocked neuropathic pain in mice.\",\n      \"method\": \"In silico docking screen, in vitro kinase assay, X-ray co-crystallography (chlortetracycline-EphB1), in vivo phosphorylation assay, neuropathic pain behavioral model\",\n      \"journal\": \"Proceedings of the National Academy of Sciences of the United States of America\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — crystal structure + in vitro kinase assay + in vivo validation, multiple orthogonal methods\",\n      \"pmids\": [\"33627480\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"EphB1 expressed in the striatal anlage signals to ephrin-B3 on migrating cortical interneurons and striatal neurons via reverse signaling with opposite effects: in cortical interneurons, EphB1-ephrin-B3 reverse signaling phosphorylates Src and FAK causing repulsion; in striatal neurons expressing ephrin-B3, EphB1 reduces pSrc and pFAK causing cessation of migration. In ephrin-B3 knockout mice, cortical interneurons are misrouted into the striatum and striatal neurons over-migrate.\",\n      \"method\": \"In vitro migration assays, ephrin-B3 knockout mouse, immunofluorescence for pSrc/pFAK, in vivo cell tracking\",\n      \"journal\": \"Frontiers in cellular neuroscience\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — in vitro assay + knockout mouse + biochemical phosphorylation in single lab\",\n      \"pmids\": [\"25100946\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2015,\n      \"finding\": \"EphB1 intracellular signaling domains are required for corpus callosum and anterior commissure formation. Truncated EphB1/EphB2 mice lacking intracellular domains show partial/complete agenesis of corpus callosum and highly penetrant anterior commissure misprojection, indicating forward (and potentially reverse) intracellular signaling by EphB1/EphB2 is required for forebrain axon guidance.\",\n      \"method\": \"Knock-in truncated EphB1/EphB2 mice, axon tract tracing, immunohistochemistry\",\n      \"journal\": \"Developmental neurobiology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — genetic knock-in with defined axon guidance phenotype, single lab\",\n      \"pmids\": [\"26148571\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2015,\n      \"finding\": \"EphB1 ligand-dependent signaling activates the DNA damage response (DDR) cascade via p53 DNA binding, leading to activation of ATR, Chk1, p53, p21, p38, CDK1(Tyr15) and Bax, and downregulation of HSP27 and Bcl2. In EphB1-methylated AML cells, re-introduction of EphB1 enhanced the DDR cascade and enforced programmed cell death, identifying EphB1 as a tumor suppressor in AML.\",\n      \"method\": \"EphB1 re-expression in methylated AML cells, signaling pathway analysis by western blot, promoter methylation analysis, p53 DNA binding assay\",\n      \"journal\": \"Molecular cancer research : MCR\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — re-expression rescue + signaling cascade analysis, single lab\",\n      \"pmids\": [\"25944917\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"Neddylation of EphB1 enhances its kinase activity by preventing its degradation, thereby promoting proliferation, migration, and activation of hepatic stellate cells. EphB1 expression and neddylation are both increased in activated hepatic stellate cells.\",\n      \"method\": \"Neddylation assay (immunoprecipitation), kinase activity assay, hepatic stellate cell functional assays (proliferation, migration)\",\n      \"journal\": \"International journal of molecular sciences\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — biochemical neddylation assay + kinase activity + functional cell assays, single lab\",\n      \"pmids\": [\"36834826\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"CRC-associated somatic mutations in EphB1 reduce kinase activity and protein stability. CRC mutant EphB1 receptors inhibit STAT3 and ERK1/2 signaling (contrasting wild-type) and are unable to suppress migration of CRC cells or compartmentalize cells in ephrin-B1 co-culture assays, establishing a kinase-dependent tumor suppressor function.\",\n      \"method\": \"Purified recombinant WT and mutant EphB1 kinase domain assays, cell migration assay, cell compartmentalization assay, mammalian cell expression\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — in vitro kinase reconstitution with purified proteins + cell-based functional assays, multiple mutants\",\n      \"pmids\": [\"37527777\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"SUMOylation of EphB1 at lysine residue 785 suppresses cell proliferation, anchorage-independent growth, and xenograft tumor growth. SUMOylated EphB1 represses activation of its downstream effector PKCγ. A reciprocal regulatory loop between PKCγ and EphB1 SUMOylation was identified.\",\n      \"method\": \"Ni2+-NTA pull-down and immunoprecipitation for SUMOylation, K785R mutant expression, soft agar colony formation, xenograft mouse model, PKCγ activation assay\",\n      \"journal\": \"Cellular physiology and biochemistry\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — biochemical SUMOylation assay + mutagenesis + in vivo tumor model, single lab\",\n      \"pmids\": [\"29550816\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"Tumor cell-expressed EPHB1 interacts with platelet-expressed EFNB1 (ephrin-B1) in the liver metastatic niche of pancreatic ductal adenocarcinoma. This contact-dependent EPHB1-EFNB1 interaction activates platelets via reverse signaling through AKT signaling; activated platelets then release serotonin (5-HT) which promotes tumor growth.\",\n      \"method\": \"Gain-of-function and loss-of-function of Ephb1, tumor cell-platelet adhesion assay, recombinant protein treatments, Tph1-knockout mice (serotonin-deficient), mCherry niche-labeling system\",\n      \"journal\": \"Cancer communications (London, England)\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — genetic LOF/GOF + knockout mice + mechanistic pathway in single lab\",\n      \"pmids\": [\"39648610\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"EphB1 in GABAergic neurons (not cortical excitatory neurons or endothelial cells) is required for proper long-range cortical glutamatergic axon guidance. Conditional EphB1 knockout in GABAergic cells (Vgat-Cre) reproduces the cortical axon guidance defects of global EphB1 KO, with misguided axon bundles containing co-mingled GABAergic and glutamatergic axons near blood vessels.\",\n      \"method\": \"Cell-type-specific conditional knockout (Vgat-Cre, Emx1-Cre, Tie2-Cre), axon tract tracing, immunohistochemistry\",\n      \"journal\": \"Development (Cambridge, England)\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — cell-type-specific conditional KO with multiple Cre lines to dissect cell-autonomous requirement, in vivo axon tracing\",\n      \"pmids\": [\"38345254\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"EphB1 in glutamatergic neurons of the ventral posteromedial thalamic nucleus (VPM) promotes emergence from anesthesia by activating the VPMGlu→primary somatosensory cortex pathway. EphrinB-EphB1 signaling excites VPMGlu neurons through NR2B phosphorylation at Tyr-1472, and disinhibits VPMGlu neurons through ubiquitin-mediated degradation of KCC2; these are identified as two independent mechanisms.\",\n      \"method\": \"Conditional knockout, in vivo electrophysiology, NR2B phosphorylation assays, KCC2 ubiquitination assay, chemogenetics/optogenetics to probe neural circuit\",\n      \"journal\": \"Science advances\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — in vivo KO + circuit-level electrophysiology + molecular mechanism, single lab\",\n      \"pmids\": [\"41348875\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"TGF-β-activated Smad2 transcriptionally upregulates endogenous EphB1 expression in lung cancer cells. Ligand-independent (unphosphorylated) EphB1 promotes epithelial-mesenchymal transition (EMT) by upregulating CDH2 (N-cadherin) and increases migration and invasion, while ligand-dependent (phosphorylated) EphB1 (activated by ephrin-B2) inhibits migration and invasion.\",\n      \"method\": \"EphB1 phosphorylation mutant expression, western blot for Smad2 and CDH2, transwell migration/invasion assay, siRNA knockdown\",\n      \"journal\": \"Journal of Cancer\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — phosphorylation mutant, siRNA, and signaling analysis, single lab\",\n      \"pmids\": [\"32368295\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"EphB1 promotes CREB phosphorylation via PI3K/AKT signaling in a cell-autonomous manner in hypothalamic neurons. EphB1 deficiency reduces hypothalamic CRH and TRH expression, leading to impaired thermogenesis and locomotion (but not food intake), and results in obesity. Intraventricular administration of TRH or CRH suppressed obesity in Ephb1 mutant mice.\",\n      \"method\": \"Forward genetic screen, hypothalamic tissue and primary cell signaling analysis, PI3K/AKT pathway inhibitors, intracerebroventricular peptide delivery, metabolic phenotyping\",\n      \"journal\": \"Obesity (Silver Spring, Md.)\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — genetic mutant + cell-autonomous signaling + pharmacological rescue, single lab\",\n      \"pmids\": [\"40207393\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"Recurring EPHB1 mutations in colorectal and other cancers alter receptor signaling and cell compartmentalization. Ligand-binding domain mutations (C61Y, R90C, R170W), fibronectin domain mutation (R351L), and kinase domain mutation (D762N) reduce compartmentalization and ligand-induced receptor phosphorylation. Kinase domain mutations R743W and G821R enhance compartmentalization without altered phosphorylation. Phosphoproteome analysis linked reduced-compartmentalization mutants to PI3K pathway/PIK3C2B phosphorylation.\",\n      \"method\": \"Confocal microscopy compartmentalization assay, phospho-proteome analysis, stable expression of 15 mutants in CRC cells\",\n      \"journal\": \"Cell communication and signaling : CCS\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — systematic functional mutagenesis panel + phosphoproteomics, single lab\",\n      \"pmids\": [\"38102712\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"EphB1 is a receptor tyrosine kinase that, upon ligand (ephrin-B) binding, undergoes autophosphorylation and recruits a set of adaptor/effector proteins (Nck via pTyr594, Grb7 via pTyr928, p52Shc/c-Src via Tyr600/778, Grb2, Cbl, PTEN/NHERF1, Porf-2/RhoGAP, Caskin1) to activate downstream JNK, ERK/MAPK, and Rac1-GTPase pathways that regulate cell migration, adhesion, and actin cytoskeletal dynamics; it controls neural circuit formation (ipsilateral retinal projection via ephrin-B2 repulsion at the optic chiasm, dopaminergic pathway specification, cortical axon guidance through GABAergic neurons), modulates synaptic plasticity and pain processing via NR2B/NMDA receptor phosphorylation, regulates caveolae biogenesis through constitutive Cav-1 interaction, and is post-translationally regulated by Cbl-mediated ubiquitination/lysosomal degradation, SUMOylation at K785 (suppressing PKCγ), and neddylation (enhancing kinase activity), with its tumor-suppressive kinase function frequently impaired by somatic mutations in colorectal and other cancers.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"EphB1 is an ephrin-B-activated receptor tyrosine kinase that, upon ligand engagement and autophosphorylation, assembles phosphotyrosine-dependent adaptor complexes to control cell adhesion, migration, and actin cytoskeletal dynamics [#0, #1, #2]. Activated EphB1 recruits the SH2-adaptor Nck through phospho-Tyr594 to drive JNK activation and cell attachment, and the Nck–paxillin/FAK axis with c-Src-dependent paxillin phosphorylation to promote migration [#1, #5]; it also engages Grb2/p52Shc and Grb7 through distinct phosphotyrosines (Tyr600/Tyr778, Tyr928) to activate ERK/MAPK and chemotaxis [#2, #3], and recruits the scaffold Caskin1 via Nck [#15]. Forward signaling brakes axon outgrowth by binding the RhoGAP Porf-2 to deactivate Rac1 [#18], while PTEN, delivered through NHERF1, antagonizes EphB1-dependent adhesion and chemotaxis [#16]. EphB1 is a principal determinant of neural circuit wiring: it functions as the preferred receptor for ephrin-B2 at the optic chiasm, where its kinase-competent forward signaling repels ventrotemporal retinal ganglion cell axons to establish the ipsilateral projection downstream of the Zic2 transcription factor [#8, #11, #12, #13], and it directs neural crest migration, dopaminergic pathway specification, and forebrain commissural and long-range cortical axon guidance—the latter cell-autonomously through GABAergic neurons [#9, #10, #14, #22, #28]. EphB1 localizes to caveolae and is required for caveolae biogenesis by constitutively binding and stabilizing caveolin-1 against ubiquitin-mediated degradation [#6, #19]. Its abundance and activity are tuned post-translationally—by Cbl-mediated ubiquitination and lysosomal degradation [#7], SUMOylation at Lys785 that suppresses PKCγ and tumor growth [#26], and neddylation that stabilizes the receptor and enhances kinase activity [#24]. Kinase-dependent EphB1 acts as a tumor suppressor: cancer-associated somatic mutations reduce its activity, stability, and ability to compartmentalize cells, and re-expression of EphB1 in silenced leukemia cells enforces a p53-driven DNA-damage/apoptotic program [#23, #25, #32].\",\n  \"teleology\": [\n    {\n      \"year\": 1998,\n      \"claim\": \"Established the first proximal EphB1 effector linkage by showing how a defined receptor phosphotyrosine couples kinase activation to a downstream MAP kinase and an adhesive cell response.\",\n      \"evidence\": \"Yeast two-hybrid, Co-IP, Y594F mutagenesis, JNK and cell attachment assays\",\n      \"pmids\": [\"9430661\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Did not define the kinase chain from Nck to JNK\", \"Physiological cell context of the attachment response not addressed\"]\n    },\n    {\n      \"year\": 1999,\n      \"claim\": \"Defined EphB1 as a ligand-density sensor whose kinase signaling, not mechanical tethering, converts ephrin-B engagement into integrin-mediated attachment.\",\n      \"evidence\": \"Reconstituted ephrin-B1 surface-density assay with integrin blocking and signaling-defective EphB1 mutants\",\n      \"pmids\": [\"10205170\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Mechanism coupling EphB1 to specific integrins not resolved\", \"Tested in endothelial and transfected HEK cells only\"]\n    },\n    {\n      \"year\": 2002,\n      \"claim\": \"Mapped a second adaptor branch by identifying Grb7 as an autophosphorylation-dependent EphB1 partner and substrate that drives motility.\",\n      \"evidence\": \"Yeast two-hybrid, Co-IP, Tyr928 mutagenesis, kinase and migration assays\",\n      \"pmids\": [\"12223469\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Downstream effectors of Grb7 in EphB1 migration unresolved\"]\n    },\n    {\n      \"year\": 2003,\n      \"claim\": \"Resolved the EphB1 migration pathway by linking Tyr600/Tyr778, c-Src, p52Shc and ERK to chemotaxis as distinct from adhesion.\",\n      \"evidence\": \"Co-IP, Y600F/Y778F mutagenesis, dominant-negative Src, PD98059/PP2 inhibitors, migration vs adhesion assays\",\n      \"pmids\": [\"12925710\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Spatial coordination of parallel adhesion and migration outputs not defined\"]\n    },\n    {\n      \"year\": 2003,\n      \"claim\": \"Demonstrated that EphB1 forward signaling drives a specific developmental axon-sorting decision, establishing its role in binocular circuit formation.\",\n      \"evidence\": \"EphB1 knockout mice, expression mapping, ephrin-B2 function-blocking in semiintact visual system\",\n      \"pmids\": [\"12971893\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Intracellular effectors mediating chiasm repulsion not identified in this study\"]\n    },\n    {\n      \"year\": 2004,\n      \"claim\": \"Built the EphB1→Nck→paxillin/FAK→Src migration complex, connecting the juxtamembrane adaptor to focal-adhesion substrate phosphorylation.\",\n      \"evidence\": \"Co-IP, paxillin Y31F/Y118F and EphB1 Y594F mutagenesis, dominant-negative Src, PP2, migration assay\",\n      \"pmids\": [\"15107421\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether the complex acts at nascent versus mature adhesions unresolved\"]\n    },\n    {\n      \"year\": 2006,\n      \"claim\": \"Placed EphB1 in caveolae and tied its membrane targeting and ERK output to a caveolin-binding motif in the kinase domain.\",\n      \"evidence\": \"Fractionation, Co-IP, Cav-1 scaffolding-domain mutant, confocal microscopy, ERK assays\",\n      \"pmids\": [\"16723736\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Did not determine whether caveolae are required for all EphB1 signaling outputs\"]\n    },\n    {\n      \"year\": 2007,\n      \"claim\": \"Defined the negative-feedback arm of EphB1 signaling through Cbl-mediated ubiquitination and lysosomal turnover.\",\n      \"evidence\": \"Co-IP, GST pull-down, Cbl ligase-dead and EphB1-K652R mutants, ubiquitination assay, bafilomycin, PP2\",\n      \"pmids\": [\"18034775\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Kinetics of receptor recycling versus degradation not quantified\"]\n    },\n    {\n      \"year\": 2008,\n      \"claim\": \"Identified the transcriptional control point upstream of EphB1 by showing Zic2 induces functional EphB1 to convert axon behavior to ephrin-B2 avoidance.\",\n      \"evidence\": \"Retinal explant electroporation, avoidance assays on ephrin-B2, growth-cone immunofluorescence\",\n      \"pmids\": [\"18524895\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Direct versus indirect regulation of the EphB1 promoter by Zic2 not established\"]\n    },\n    {\n      \"year\": 2009,\n      \"claim\": \"Showed EphB1 is both necessary and sufficient for ipsilateral redirection and that its specificity resides in extracellular and juxtamembrane domains.\",\n      \"evidence\": \"In utero retinal electroporation, EphB1-EphB2 chimeric receptors, in vivo tracing\",\n      \"pmids\": [\"19295152\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Molecular basis of domain-specific specificity over EphB2 not defined at residue level\"]\n    },\n    {\n      \"year\": 2011,\n      \"claim\": \"Distinguished forward from reverse signaling, proving EphB1 intracellular signaling drives the ipsilateral projection.\",\n      \"evidence\": \"Intracellular-truncated EphB1 knock-in mice, axon tracing, EphB/ephrin-B genetic combinations\",\n      \"pmids\": [\"22103419\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Effector cascade downstream of forward signal in RGCs not fully mapped\"]\n    },\n    {\n      \"year\": 2012,\n      \"claim\": \"Extended EphB1 adaptor wiring by identifying Caskin1 as a Nck-bridged substrate undergoing phosphorylation-induced conformational change.\",\n      \"evidence\": \"Co-IP, mass-spectrometry phosphosite mapping, CD spectroscopy\",\n      \"pmids\": [\"23181695\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Functional consequence of Caskin1 phosphorylation for EphB1 output untested\"]\n    },\n    {\n      \"year\": 2012,\n      \"claim\": \"Revealed PTEN/NHERF1 as a phosphatase brake that disrupts the PTEN-Cbl complex and limits EphB1-driven adhesion and chemotaxis.\",\n      \"evidence\": \"Co-IP, PTEN and NHERF1 siRNA, PTEN phosphatase mutants, adhesion and chemotaxis assays\",\n      \"pmids\": [\"23118026\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"In vivo relevance of the PTEN-Cbl switch not addressed\"]\n    },\n    {\n      \"year\": 2018,\n      \"claim\": \"Identified the RhoGAP Porf-2 as the link between EphB1 forward signaling and Rac1 inactivation in axon-growth braking.\",\n      \"evidence\": \"Co-IP, GTP-Rac1 pull-down, GAP-domain mutagenesis, axon growth measurement\",\n      \"pmids\": [\"29938386\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Recruitment mechanism of Porf-2 to activated EphB1 unresolved\"]\n    },\n    {\n      \"year\": 2018,\n      \"claim\": \"Established SUMOylation at Lys785 as a tumor-suppressive post-translational switch that represses PKCγ.\",\n      \"evidence\": \"Ni-NTA/IP SUMOylation assay, K785R mutant, soft-agar and xenograft assays, PKCγ activation assay\",\n      \"pmids\": [\"29550816\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"SUMO ligase responsible not identified\", \"Relationship to kinase activity not defined\"]\n    },\n    {\n      \"year\": 2020,\n      \"claim\": \"Demonstrated that EphB1 constitutively stabilizes caveolin-1 and is required for caveolae biogenesis in endothelial cells.\",\n      \"evidence\": \"Super-resolution microscopy, FRET, Co-IP, CSD-motif deletion mutant, EphB1-KO ECs, EM caveolae counting\",\n      \"pmids\": [\"32238105\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Physiological vascular consequences of caveolae loss not assessed\"]\n    },\n    {\n      \"year\": 2020,\n      \"claim\": \"Uncovered phosphorylation-state-dependent dual function: ligand-independent EphB1 promotes EMT while activated EphB1 inhibits invasion, downstream of TGF-β/Smad2.\",\n      \"evidence\": \"EphB1 phosphorylation mutants, Smad2/CDH2 western blot, transwell assays, siRNA\",\n      \"pmids\": [\"32368295\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Mechanism by which unphosphorylated EphB1 upregulates N-cadherin unclear\"]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"Provided a structural and pharmacological handle by co-crystallizing a tetracycline in the EphB1 ATP pocket and blocking neuropathic pain in vivo.\",\n      \"evidence\": \"In silico docking, in vitro kinase assay, X-ray co-crystallography, in vivo phosphorylation and pain model\",\n      \"pmids\": [\"33627480\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Selectivity over related Eph kinases not fully characterized\"]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Established EphB1 kinase activity as essential for its tumor-suppressor function, with cancer mutations crippling activity, stability and cell compartmentalization.\",\n      \"evidence\": \"Purified WT/mutant kinase-domain assays, migration and compartmentalization assays in CRC cells\",\n      \"pmids\": [\"37527777\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"In vivo tumor-suppressor validation of specific mutants not shown\"]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Showed neddylation stabilizes EphB1 and enhances kinase activity to promote hepatic stellate cell activation.\",\n      \"evidence\": \"Neddylation IP, kinase activity assay, proliferation and migration assays\",\n      \"pmids\": [\"36834826\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Neddylation site and responsible ligase not mapped\"]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Systematically parsed cancer-mutation effects, linking reduced-compartmentalization mutants to altered PI3K-pathway phosphorylation.\",\n      \"evidence\": \"Confocal compartmentalization assay and phosphoproteomics across 15 mutants in CRC cells\",\n      \"pmids\": [\"38102712\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Causal role of PIK3C2B phosphorylation in compartmentalization not tested\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Pinpointed the cell-autonomous requirement for EphB1 in GABAergic neurons for long-range cortical glutamatergic axon guidance.\",\n      \"evidence\": \"Cell-type-specific conditional knockout (Vgat-Cre, Emx1-Cre, Tie2-Cre), axon tracing, IHC\",\n      \"pmids\": [\"38345254\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Molecular signal between GABAergic EphB1 and glutamatergic axons not defined\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Revealed a tumor-platelet axis in which tumor EPHB1 activates platelets via ephrin-B1 reverse signaling to release pro-tumor serotonin.\",\n      \"evidence\": \"Ephb1 GOF/LOF, tumor-platelet adhesion assay, recombinant proteins, Tph1-knockout mice, niche labeling\",\n      \"pmids\": [\"39648610\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Generalizability beyond pancreatic liver-metastatic niche untested\"]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Defined dual molecular mechanisms—NR2B Tyr1472 phosphorylation and KCC2 degradation—by which EphB1 drives thalamocortical excitation and anesthesia emergence.\",\n      \"evidence\": \"Conditional KO, in vivo electrophysiology, NR2B phosphorylation and KCC2 ubiquitination assays, chemo/optogenetics\",\n      \"pmids\": [\"41348875\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Relative contribution of the two mechanisms in vivo not quantified\"]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Linked EphB1 to hypothalamic energy homeostasis via cell-autonomous PI3K/AKT/CREB control of CRH/TRH and thermogenesis.\",\n      \"evidence\": \"Forward genetic screen, PI3K/AKT inhibitors, ICV peptide rescue, metabolic phenotyping\",\n      \"pmids\": [\"40207393\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Whether the phenotype requires ligand engagement or kinase activity unresolved\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"How EphB1's many context-dependent outputs—forward vs reverse signaling, kinase-dependent tumor suppression vs ligand-independent EMT promotion, and tissue-specific effector selection—are integrated at a structural and regulatory level remains unresolved.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No unified model linking post-translational modifications (Cbl, SUMO, neddylation) to effector choice\", \"Determinants of receptor specificity over other EphB receptors in vivo undefined\", \"Structural basis of compartmentalization versus signaling outputs unknown\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0140096\", \"supporting_discovery_ids\": [1, 2, 3, 5, 25]},\n      {\"term_id\": \"GO:0016740\", \"supporting_discovery_ids\": [3, 20, 25]},\n      {\"term_id\": \"GO:0060089\", \"supporting_discovery_ids\": [0, 8, 13]},\n      {\"term_id\": \"GO:0060090\", \"supporting_discovery_ids\": [1, 15]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005886\", \"supporting_discovery_ids\": [6, 19]},\n      {\"term_id\": \"GO:0031410\", \"supporting_discovery_ids\": [6, 19]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-162582\", \"supporting_discovery_ids\": [1, 2, 5]},\n      {\"term_id\": \"R-HSA-1266738\", \"supporting_discovery_ids\": [8, 12, 13, 28]},\n      {\"term_id\": \"R-HSA-1643685\", \"supporting_discovery_ids\": [23, 25, 32]},\n      {\"term_id\": \"R-HSA-112316\", \"supporting_discovery_ids\": [29, 31]}\n    ],\n    \"complexes\": [\"caveolae\"],\n    \"partners\": [\"EFNB2\", \"EFNB1\", \"NCK1\", \"GRB7\", \"SHC1\", \"SRC\", \"CBL\", \"CAV1\"],\n    \"other_free_text\": []\n  }\n}","audit_flag":null,"evaluation":{"pairwise":"win","faith_supported":7,"faith_total":7,"faith_pct":100.0}}