{"gene":"IRF6","run_date":"2026-06-10T01:55:23","timeline":{"discoveries":[{"year":2002,"finding":"Loss-of-function (nonsense/missense) mutations in IRF6 cause Van der Woude syndrome and popliteal pterygium syndrome. IRF6 mRNA is highly expressed along the medial edge of the fusing palate, tooth buds, hair follicles, genitalia and skin, establishing a direct role for IRF6 in orofacial and epithelial development.","method":"Mutation screening of VWS/PPS families; in situ expression analysis of Irf6 mRNA","journal":"Nature genetics","confidence":"High","confidence_rationale":"Tier 2 / Strong — direct mutation identification in >45 families plus expression analyses; replicated across multiple independent cohorts in subsequent studies","pmids":["12219090"],"is_preprint":false},{"year":2006,"finding":"Mice homozygous for a missense mutation in Irf6 develop a hyperproliferative epidermis that fails to undergo terminal differentiation, resulting in soft tissue fusions. Compound heterozygosity for Irf6 and Sfn (14-3-3σ/stratifin) recapitulates similar keratinizing epithelial defects, placing Irf6 and Sfn in a shared genetic pathway controlling the keratinocyte proliferation-differentiation switch.","method":"Mouse genetics (homozygous missense knock-in); compound heterozygote epistasis analysis","journal":"Nature genetics","confidence":"High","confidence_rationale":"Tier 2 / Strong — genetic epistasis with defined cellular phenotype (hyperproliferation/failure of terminal differentiation); compound heterozygote experiment provides independent mechanistic validation","pmids":["17041603"],"is_preprint":false},{"year":2009,"finding":"IRF6 is essential for oral epithelial differentiation and functions in a convergent molecular pathway with the Notch ligand Jagged2. IRF6 plays a key role in formation and maintenance of the oral periderm, with spatio-temporal regulation of this periderm being essential for appropriate palatal adhesion and fusion.","method":"Mouse genetic analysis (Irf6 mutant and Jagged2 mutant); histological and molecular characterization of palatal/oral epithelium","journal":"Human molecular genetics","confidence":"High","confidence_rationale":"Tier 2 / Strong — genetic epistasis between Irf6 and Jagged2, with mechanistic characterization of periderm formation and palatal adhesion phenotype","pmids":["19439425"],"is_preprint":false},{"year":2011,"finding":"IRF6 is transcriptionally activated by p63 and, in turn, induces proteasome-mediated down-regulation of p63, forming a regulatory feedback loop that limits keratinocyte proliferative potential. ChIP-seq and gene expression profiling after siRNA-mediated IRF6 depletion revealed direct IRF6 target genes involved in cell cycle, differentiation, cell adhesion, and cell–cell contact. IRF6 down-regulation promotes invasive behavior in squamous cell carcinoma cells.","method":"ChIP-seq for IRF6 binding sites; siRNA knockdown; gene expression profiling; in vitro invasion assays; re-expression of IRF6 in SCC cells","journal":"Proceedings of the National Academy of Sciences of the United States of America","confidence":"High","confidence_rationale":"Tier 2 / Strong — multiple orthogonal methods (ChIP-seq, siRNA KD, invasion assay, reintroduction rescue); single lab but mechanistically comprehensive","pmids":["21807998"],"is_preprint":false},{"year":2011,"finding":"IRF6 is induced during keratinocyte differentiation through a Notch-dependent mechanism and is a primary Notch target gene in keratinocytes and keratinocyte-derived SCC cells. Increased IRF6 expression contributes to Notch-driven regulation of growth/differentiation genes, while it is not required for induction of canonical Notch targets (p21WAF1/Cip1, Hes1, Hey1). Down-modulation of IRF6 counteracts differentiation and promotes ras-induced tumour formation.","method":"Notch gain-of-function; IRF6 siRNA knockdown; gene expression analysis; in vivo tumour formation assay","journal":"The EMBO journal","confidence":"High","confidence_rationale":"Tier 2 / Strong — multiple orthogonal approaches (gain-of-function, knockdown, in vivo tumour model), distinct from but complementary to PMID 21807998","pmids":["21909072"],"is_preprint":false},{"year":2013,"finding":"TGFβ signaling regulates expression of Irf6, and IRF6 and SMAD4 synergistically regulate the fate of the medial edge epithelium (MEE) during palatal fusion. Haploinsufficiency of Irf6 in Smad4 conditional knockout mice causes compromised p21 expression and MEE persistence. Overexpression of Irf6 rescued p21 expression and MEE degeneration in Tgfbr2-deficient mice, establishing TGFβ-mediated Irf6 activity as responsible for MEE degeneration.","method":"Mouse genetics (conditional knockout, compound heterozygotes); Irf6 overexpression rescue experiment; p21 immunostaining","journal":"Development (Cambridge, England)","confidence":"High","confidence_rationale":"Tier 2 / Strong — genetic epistasis with rescue experiment using Irf6 overexpression; multiple mutant combinations tested","pmids":["23406900"],"is_preprint":false},{"year":2019,"finding":"RIPK4 kinase activity is required for mouse development; RIPK4 phosphorylates IRF6 at Ser413 and Ser424, priming IRF6 for activation. RNA-seq, ChIP-seq, and ATAC-seq of wild-type versus IRF6-deficient skin showed IRF6 is enriched at bivalent promoters and its deficiency causes defective expression of genes involved in lipid metabolism and tight junction formation, culminating in a severe epidermal barrier defect.","method":"Kinase-dead knock-in mouse; phosphosite mapping (Ser413/Ser424); RNA-seq; ChIP-seq; ATAC-seq; lipid composition analysis of stratum corneum","journal":"Nature","confidence":"High","confidence_rationale":"Tier 1–2 / Strong — phosphorylation site identified with mutagenesis context, combined with genome-wide chromatin and transcriptomic profiling and functional barrier assay; highly rigorous single study","pmids":["31578523"],"is_preprint":false},{"year":2015,"finding":"TLR3 stimulation of keratinocytes upregulates IRF6, which selectively promotes IL-23p19 expression while suppressing IFN-β expression. IRF6 silencing enhanced poly(IC)-inducible IFN-β mRNA and inhibited IL-23p19 mRNA. Co-transfection of IRF6 increased IL-23p19 promoter activity but inhibited IFN-β promoter activity, defining IRF6 as a specificity-determining downstream mediator of TLR3 signalling.","method":"siRNA silencing of IRF6; gene reporter (promoter) assays; co-immunoprecipitation; proximity ligation assay","journal":"Immunology and cell biology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — siRNA knockdown plus reporter assays in primary keratinocytes; single lab with two orthogonal approaches","pmids":["26303210"],"is_preprint":false},{"year":2016,"finding":"IRF6 acts downstream of IRAK1 to stimulate expression of the pro-inflammatory cytokine IL-36γ in oral epithelial cells in response to Porphyromonas gingivalis (via TLR2). Gene silencing and promoter reporter experiments confirmed IRF6 drives IL-36γ transcription.","method":"Gene silencing (siRNA); promoter reporter assays; TLR2 agonist stimulation","journal":"Journal of immunology (Baltimore, Md. : 1950)","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — siRNA plus promoter assays; single lab, two orthogonal approaches","pmids":["26819203"],"is_preprint":false},{"year":2016,"finding":"RIPK4 overexpression in oral keratinocytes induces CCL5 and CXCL11 expression in an IRF6-dependent manner. Gene silencing of RIPK4 or IRF6 abolished PKC pathway-induced CCL5/CXCL11 expression. Promoter reporter assays showed RIPK4 stimulates transactivation of CCL5 and CXCL11 promoters by IRF6, establishing a RIPK4→IRF6 signalling axis for proinflammatory cytokine induction in keratinocytes.","method":"RIPK4 overexpression; siRNA knockdown of RIPK4 and IRF6; gene reporter (promoter) assays; PKC pathway activation","journal":"Cytokine","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — overexpression, siRNA knockdown, and promoter reporter assays; single lab with multiple orthogonal methods","pmids":["27014863"],"is_preprint":false},{"year":2015,"finding":"IRF6 is a mediator of TGFβ3 during palatal fusion: ectopic IRF6 expression rescued shTgfβ3-induced fusion defects in palatal organ culture. IRF6 increases SNAI2 (Slug) expression and decreases epithelial markers (E-cadherin, Plakophilin, ZO-1), regulating epithelial–mesenchymal transition (EMT) during palatal fusion. Knockdown of Snai2 abolished the IRF6 rescuing effect.","method":"Palatal shelf organ culture; shRNA knockdown; ectopic IRF6 expression; RT-PCR and protein analysis of EMT markers","journal":"Scientific reports","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — organ culture rescue experiments with knockdown and overexpression; single lab, multiple approaches","pmids":["26240017"],"is_preprint":false},{"year":2017,"finding":"IRF6 suppresses PPARγ transcription by binding IRF recognition sites upstream of the PPARγ coding region, thereby suppressing alternative (M2) macrophage polarization. ChIP confirmed IRF6 occupancy at the PPARγ locus. IRF6 knockdown enhanced M2 activation; IRF6 overexpression dramatically attenuated it.","method":"Gain- and loss-of-function (overexpression and siRNA); chromatin immunoprecipitation (ChIP); computational binding site prediction","journal":"Endocrinology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — ChIP plus gain/loss-of-function; single lab with orthogonal methods","pmids":["28645193"],"is_preprint":false},{"year":2014,"finding":"A regulatory mutation (350dupA) in the IRF6 enhancer element MCS9.7 abrogates binding of p63 and E47 transcription factors to overlapping cis-motifs, significantly disrupting enhancer activity. The 350dupA simultaneously creates a CAAAGT motif bound by Lef1; Lef1/β-Catenin chimeric protein repressed MCS9.7-350dupA enhancer activity, demonstrating a dual loss-of-function/gain-of-function mechanism regulating IRF6 expression.","method":"Enhancer reporter assays in human cell cultures; transgenic mouse lacZ reporter assay; site-directed mutagenesis of binding motifs; Lef1 overexpression; electrophoretic mobility shift / binding assays","journal":"Human molecular genetics","confidence":"High","confidence_rationale":"Tier 1–2 / Strong — multiple orthogonal methods (cell reporter assay, transgenic mouse assay, mutagenesis, protein binding disruption) in a single rigorous study","pmids":["24442519"],"is_preprint":false},{"year":2017,"finding":"IRF6 is identified as a direct protein interactor of NME1 and NME2 (regulators of Rho-type GTPases and E-cadherin endocytosis) by yeast two-hybrid screening and co-immunoprecipitation. The IRF6–NME interaction is enhanced by phosphorylation of key serine residues in the IRF6 C-terminus. CLP-associated IRF6 missense mutations disrupt NME binding and result in elevated Rac1 and RhoA activation. NME1 and NME2 co-localize with IRF6 in the cytoplasm of primary palatal epithelial cells in vivo.","method":"Yeast two-hybrid screen; co-immunoprecipitation; phosphorylation assays; Rac1/RhoA activation assays; co-localization in primary palatal epithelial cells","journal":"Journal of dental research","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — yeast 2-hybrid plus co-IP plus functional Rho GTPase readout; single lab with multiple orthogonal methods","pmids":["28767310"],"is_preprint":false},{"year":2019,"finding":"Overexpression of Irf6 in mice causes exencephaly through suppression of Tfap2a and Grhl3 expression, while loss of Irf6 function causes curly tail with reduced Tfap2a and Grhl3 expression in tail tissues, demonstrating that a Tfap2a–Irf6–Grhl3 genetic pathway operates during neurulation as well as orofacial development.","method":"Transgenic Irf6 overexpression mouse model; Irf6 loss-of-function mouse; gene expression analysis of Tfap2a and Grhl3","journal":"Human molecular genetics","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — genetic gain- and loss-of-function with molecular readout of pathway targets; single lab","pmids":["30689861"],"is_preprint":false},{"year":2017,"finding":"Loss of Irf6 in mice causes craniosynostosis and mandibular hypoplasia. Double heterozygous Irf6+/−;Twist1+/− embryos show severe mandibular hypoplasia with reduced EDN1, DLX5, DLX6 and HAND2 expression in mesenchymal cells, demonstrating an intercellular genetic interaction between Irf6 (expressed in epithelium) and Twist1 (expressed in mesenchyme). Exogenous EDN1 peptide partially rescued Meckel's cartilage abnormalities.","method":"Mouse genetics (single and double heterozygotes); gene expression analysis; mandibular explant culture with EDN1 rescue","journal":"Scientific reports","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — genetic epistasis with molecular readout and partial rescue; single lab","pmids":["28769044"],"is_preprint":false},{"year":2013,"finding":"Irf6 is expressed in Myf5+ (segmental paraxial mesoderm) cells of the developing tongue. Loss of Irf6 causes reduced and poorly organized Myf5+ cell lineage and aberrant cytoskeletal formation. Non-cell-autonomously, loss of Irf6 alters Bmp2, Bmp4, Shh, and Fgf10 signalling in the tongue, establishing both cell-autonomous (muscle differentiation/cytoskeletal organization) and non-cell-autonomous (cranial neural crest-derived inter-molar eminence) roles for IRF6.","method":"Lineage tracing (fate mapping); dual-label immunostaining; Irf6 knockout mouse analysis; molecular analysis of signalling pathway genes","journal":"PloS one","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — lineage tracing combined with KO analysis and molecular pathway readout; single lab","pmids":["23451037"],"is_preprint":false},{"year":2020,"finding":"In zebrafish, irf6 regulates the expression of esrp1. Genetic disruption of irf6 and esrp1/2 in zebrafish results in cleft of the anterior neurocranium due to impaired chondrogenesis (not a neural crest migration defect). Mouse compound mutant analysis showed genetic interaction between Irf6 and Esrp1, with Irf6 heterozygosity reducing Esrp1/2 cleft severity, defining an Irf6–Esrp1/2 regulatory axis in midface morphogenesis.","method":"Zebrafish genetic knockouts; cranial neural crest lineage tracing; mouse compound mutant breeding; gene expression analysis","journal":"Development (Cambridge, England)","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — genetic epistasis across two species (zebrafish and mouse) with lineage tracing; single lab","pmids":["33234718"],"is_preprint":false},{"year":2020,"finding":"SPECC1L expression is drastically reduced in Irf6 mutant palatal shelves, placing SPECC1L downstream of IRF6 in palatogenesis. Specc1l mutant mice display periderm layer abnormalities and transient oral epithelial adhesions similar to Irf6 hypomorphic mutants.","method":"Irf6 mutant mouse analysis; immunostaining for SPECC1L expression; phenotypic comparison of mutant palatal shelves","journal":"Human molecular genetics","confidence":"Medium","confidence_rationale":"Tier 2 / Weak — loss-of-function with molecular readout showing pathway placement; single lab, single method for the IRF6-SPECC1L relationship","pmids":["31943082"],"is_preprint":false},{"year":2024,"finding":"In a mouse PDAC model, acquired resistance to immunotherapy is associated with EMT. EMT-transcription factors ZEB1 and SNAIL epigenetically and transcriptionally silence IRF6, rendering tumor cells resistant to the pro-apoptotic effects of TNF-α and insensitive to T-cell killing. This identifies IRF6 as a downstream mediator of EMT-driven immune evasion.","method":"Mouse PDAC tumor relapse model; EMT transcription factor manipulation; epigenetic analysis; TNF-α killing assays","journal":"Nature communications","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — in vivo tumor model plus mechanistic TNF-α sensitivity assay linking IRF6 silencing to apoptotic resistance; single lab","pmids":["38378697"],"is_preprint":false},{"year":2008,"finding":"IRF6 interacts with maspin (a known tumor suppressor) in mammary epithelial cells and functions to regulate cell cycle exit and entry into G0 quiescence, promoting mammary epithelial cell differentiation.","method":"Protein interaction (co-immunoprecipitation implied by 'interaction with maspin'); cell cycle analysis in mammary epithelial cells","journal":"Cell cycle (Georgetown, Tex.)","confidence":"Low","confidence_rationale":"Tier 3 / Weak — review/perspective piece describing prior interaction finding without detailed experimental reporting in this abstract; single method implied","pmids":["18604160"],"is_preprint":false},{"year":2020,"finding":"IRF6 inhibits transcription of PKM2 and GLUT1 in glioma cells, thereby impairing glycolysis. SNHG14 lncRNA sequesters STAU1 to prevent IRF6 mRNA degradation via its 3' UTR, increasing IRF6 expression; Lin28A stabilizes SNHG14. Depleting Lin28A or SNHG14 or overexpressing IRF6 reduced xenograft tumor growth.","method":"siRNA/shRNA knockdown; overexpression; ChIP (IRF6 binding to PKM2/GLUT1 promoters); xenograft mouse model; RT-PCR and western blot","journal":"Cell death & disease","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — ChIP plus gain/loss-of-function plus in vivo xenograft; single lab with multiple orthogonal methods","pmids":["32527996"],"is_preprint":false}],"current_model":"IRF6 is a transcription factor that occupies a central node in epithelial differentiation: it is transcriptionally activated by p63 (via the MCS9.7 enhancer) and feeds back to suppress p63 via proteasomal degradation; it is phosphorylated at Ser413/Ser424 by the kinase RIPK4, which primes its transcriptional activity; it regulates a broad program of keratinocyte terminal differentiation (including lipid metabolism and tight junction genes), controls the proliferation-to-differentiation switch, operates downstream of TGFβ/SMAD4 and Notch signalling in palatal medial edge epithelium, interacts genetically with Jagged2, Sfn, Esrp1/2, Twist1, and GRHL3 during craniofacial and neural tube morphogenesis, drives IL-36γ and CCL5/CXCL11 expression downstream of TLR2/3 and RIPK4 in keratinocytes, suppresses PPARγ to limit M2 macrophage polarization, and is epigenetically silenced by EMT transcription factors to render tumor cells resistant to TNF-α-mediated killing."},"narrative":{"mechanistic_narrative":"IRF6 is a transcription factor that occupies a central node in epithelial differentiation and craniofacial morphogenesis, where it governs the keratinocyte proliferation-to-differentiation switch and the loss-of-function mutations cause the Mendelian orofacial clefting disorders Van der Woude syndrome and popliteal pterygium syndrome [PMID:12219090]. IRF6 sits within a regulatory circuit in which it is transcriptionally activated by p63 through the MCS9.7 enhancer and reciprocally drives proteasomal down-regulation of p63, a feedback loop that limits keratinocyte proliferative potential and restrains invasive behavior [PMID:21807998, PMID:24442519]; it is also a primary Notch target gene whose induction promotes terminal differentiation and opposes ras-driven tumorigenesis [PMID:21909072]. Full transcriptional activation requires priming by the kinase RIPK4, which phosphorylates IRF6 at Ser413 and Ser424; through this axis IRF6 occupies bivalent promoters and directs a terminal-differentiation program including lipid metabolism and tight junction genes necessary for the epidermal barrier [PMID:31578523]. During palatal fusion IRF6 acts downstream of TGFβ/SMAD4 signalling to drive p21 expression and medial edge epithelium degeneration [PMID:23406900], operates in convergent pathways with the Notch ligand Jagged2 and the stratifin Sfn to maintain the oral periderm [PMID:17041603, PMID:19439425], and participates in genetic networks with Twist1, Esrp1/2, Tfap2a–Grhl3, and SPECC1L that pattern craniofacial and neural tube development [PMID:30689861, PMID:28769044, PMID:33234718, PMID:31943082]. Beyond development, IRF6 functions as a specificity-determining mediator of innate immune signalling in keratinocytes, driving IL-36γ, CCL5/CXCL11, and IL-23p19 expression downstream of TLR2/TLR3 and RIPK4 [PMID:26303210, PMID:26819203, PMID:27014863], and it suppresses PPARγ to constrain M2 macrophage polarization [PMID:28645193]. In cancer, epigenetic silencing of IRF6 by the EMT factors ZEB1 and SNAIL confers resistance to TNF-α-mediated apoptosis and T-cell killing [PMID:38378697].","teleology":[{"year":2002,"claim":"Establishing that IRF6 mutations cause human orofacial clefting syndromes defined IRF6 as an essential regulator of epithelial and craniofacial development, anchoring all subsequent mechanistic work.","evidence":"Mutation screening of Van der Woude/popliteal pterygium syndrome families plus in situ expression of Irf6 mRNA","pmids":["12219090"],"confidence":"High","gaps":["Molecular target genes of IRF6 not yet defined","Upstream regulators of IRF6 expression unknown"]},{"year":2006,"claim":"Mouse genetics showed that IRF6 controls the keratinocyte proliferation-to-differentiation switch, explaining the epithelial fusions seen in disease.","evidence":"Homozygous missense knock-in mouse and Irf6;Sfn compound heterozygote epistasis with hyperproliferation/differentiation-failure phenotype","pmids":["17041603"],"confidence":"High","gaps":["Molecular mechanism by which IRF6 enforces differentiation not resolved","How Sfn and IRF6 intersect biochemically unknown"]},{"year":2009,"claim":"IRF6 was placed in a convergent pathway with the Notch ligand Jagged2 controlling oral periderm formation, linking it to palatal adhesion and fusion.","evidence":"Mouse genetic analysis of Irf6 and Jagged2 mutants with histological characterization of periderm","pmids":["19439425"],"confidence":"High","gaps":["Direct biochemical link between IRF6 and Notch signalling not established here","Periderm-specific IRF6 targets unidentified"]},{"year":2011,"claim":"Defining the p63→IRF6→p63 feedback loop and genome-wide IRF6 targets converted IRF6 from a genetic locus into a characterized transcription factor restraining proliferation and invasion.","evidence":"ChIP-seq, siRNA knockdown, gene expression profiling, invasion assays and IRF6 re-expression in SCC cells; plus Notch gain/loss-of-function and in vivo tumour assays","pmids":["21807998","21909072"],"confidence":"High","gaps":["Mechanism of IRF6-driven p63 proteasomal degradation not detailed","Post-translational control of IRF6 activity not yet known"]},{"year":2013,"claim":"Genetic and rescue experiments established TGFβ/SMAD4 as an upstream activator of IRF6 driving p21 expression and medial edge epithelium degeneration during palatal fusion.","evidence":"Mouse conditional knockouts, compound heterozygotes, and Irf6 overexpression rescue of Tgfbr2-deficient MEE persistence with p21 immunostaining","pmids":["23406900"],"confidence":"High","gaps":["Direct transcriptional regulation of Irf6 by SMAD4 not demonstrated","Whether p21 is a direct IRF6 target unresolved"]},{"year":2013,"claim":"Lineage tracing revealed both cell-autonomous (muscle/cytoskeletal) and non-cell-autonomous roles for IRF6 in tongue development, broadening its developmental scope beyond epithelium.","evidence":"Fate mapping, Irf6 knockout analysis, and molecular readout of Bmp/Shh/Fgf signalling in tongue","pmids":["23451037"],"confidence":"Medium","gaps":["Mechanism of non-cell-autonomous signalling effects unknown","Single lab, correlative pathway readouts"]},{"year":2014,"claim":"Dissection of the MCS9.7 enhancer showed how a regulatory mutation reprograms IRF6 expression through dual loss and gain of transcription-factor binding, explaining noncoding clefting risk.","evidence":"Enhancer reporter assays, transgenic mouse lacZ assay, site-directed mutagenesis, and binding assays for p63/E47/Lef1","pmids":["24442519"],"confidence":"High","gaps":["In vivo contribution of Lef1/β-Catenin repression not quantified","Other enhancers controlling IRF6 not mapped"]},{"year":2015,"claim":"Functional studies positioned IRF6 as a downstream effector of TGFβ3-driven EMT during palatal fusion, acting through SNAI2.","evidence":"Palatal organ culture rescue, shRNA knockdown, ectopic IRF6 expression, and EMT marker analysis","pmids":["26240017"],"confidence":"Medium","gaps":["Whether SNAI2 is a direct IRF6 target not shown","Reconciliation with IRF6's anti-EMT role in tumors unresolved"]},{"year":2015,"claim":"IRF6 was identified as a specificity-determining switch in TLR3 signalling, biasing keratinocytes toward IL-23p19 and away from IFN-β.","evidence":"siRNA silencing, promoter reporter assays, co-IP and proximity ligation in primary keratinocytes","pmids":["26303210"],"confidence":"Medium","gaps":["IRF6 binding partner mediating promoter selectivity not defined","In vivo relevance to skin immunity not tested"]},{"year":2016,"claim":"Two studies extended IRF6 into innate immune cytokine induction, defining IRAK1→IRF6 (IL-36γ) and RIPK4→IRF6 (CCL5/CXCL11) axes in epithelial inflammation.","evidence":"siRNA silencing and promoter reporter assays with TLR2/PKC stimulation and RIPK4 overexpression in oral keratinocytes","pmids":["26819203","27014863"],"confidence":"Medium","gaps":["Direct IRF6 binding to cytokine promoters not all confirmed by ChIP","Single lab reporter-based evidence"]},{"year":2017,"claim":"Identification of NME1/NME2 as direct IRF6 interactors revealed a cytoplasmic, phosphorylation-dependent function linking IRF6 to Rho GTPase regulation, beyond its nuclear transcriptional role.","evidence":"Yeast two-hybrid, co-IP, phosphorylation and Rac1/RhoA activation assays, and co-localization in primary palatal epithelial cells","pmids":["28767310"],"confidence":"Medium","gaps":["Functional consequence of cytoplasmic IRF6 in vivo unclear","How nuclear vs cytoplasmic IRF6 pools are partitioned unknown"]},{"year":2017,"claim":"Epistasis with Twist1 demonstrated an epithelial-to-mesenchymal intercellular signalling role for IRF6 acting through EDN1/DLX/HAND2 in craniofacial skeletal development.","evidence":"Mouse single/double heterozygotes, gene expression analysis, and EDN1 rescue of mandibular explants","pmids":["28769044"],"confidence":"Medium","gaps":["Secreted mediator transmitting the epithelial-mesenchymal signal not identified","Direct vs indirect regulation of EDN1 unknown"]},{"year":2017,"claim":"IRF6 was shown to directly repress PPARγ transcription, constraining M2 macrophage polarization and extending IRF6 function to immune cell programming.","evidence":"Gain/loss-of-function and ChIP confirming IRF6 occupancy at the PPARγ locus","pmids":["28645193"],"confidence":"Medium","gaps":["Physiological context of macrophage IRF6 expression unclear","Single lab"]},{"year":2019,"claim":"Identification of RIPK4 as the kinase phosphorylating IRF6 at Ser413/Ser424 provided the priming mechanism for IRF6 activation and tied IRF6 directly to bivalent-promoter control of the epidermal barrier program.","evidence":"Kinase-dead knock-in mouse, phosphosite mapping, RNA-seq/ChIP-seq/ATAC-seq, and stratum corneum lipid/barrier assays","pmids":["31578523"],"confidence":"High","gaps":["Structural basis of phospho-priming not resolved","How phosphorylation alters IRF6 DNA binding/coactivator recruitment unknown"]},{"year":2019,"claim":"Gain- and loss-of-function established a Tfap2a–Irf6–Grhl3 genetic pathway operating in neurulation, generalizing IRF6's morphogenetic role to neural tube closure.","evidence":"Transgenic Irf6 overexpression and loss-of-function mice with Tfap2a/Grhl3 expression analysis","pmids":["30689861"],"confidence":"Medium","gaps":["Direct transcriptional regulation among pathway members not demonstrated","Single lab"]},{"year":2020,"claim":"Cross-species genetics defined an Irf6–Esrp1/2 axis and placed SPECC1L downstream of IRF6 in midface and periderm morphogenesis, refining the developmental network.","evidence":"Zebrafish knockouts with neural crest lineage tracing, mouse compound mutants, and Irf6-mutant SPECC1L immunostaining","pmids":["33234718","31943082"],"confidence":"Medium","gaps":["Direct vs indirect regulation of esrp1/SPECC1L by IRF6 not established","Single method for the IRF6-SPECC1L link"]},{"year":2020,"claim":"IRF6 was shown to repress glycolytic genes PKM2 and GLUT1 in glioma and to be controlled post-transcriptionally by a Lin28A/SNHG14/STAU1 axis, extending IRF6 to tumor metabolism.","evidence":"ChIP, knockdown/overexpression, and xenograft assays","pmids":["32527996"],"confidence":"Medium","gaps":["Relevance of glioma metabolic role to epithelial IRF6 biology unclear","Single lab"]},{"year":2024,"claim":"IRF6 was identified as a downstream mediator of EMT-driven immune evasion, with ZEB1/SNAIL silencing of IRF6 conferring resistance to TNF-α apoptosis and T-cell killing.","evidence":"Mouse PDAC relapse model, EMT-TF manipulation, epigenetic analysis, and TNF-α killing assays","pmids":["38378697"],"confidence":"Medium","gaps":["IRF6 target genes mediating TNF-α sensitivity not defined","Whether human tumors recapitulate this silencing not shown"]},{"year":null,"claim":"The structural and biochemical basis by which phosphorylation, cytoplasmic interactors, and chromatin context switch IRF6 between activator and repressor functions across epithelial, immune, and tumor contexts remains unresolved.","evidence":"","pmids":[],"confidence":"Medium","gaps":["No structural model of activated IRF6 on target promoters","Determinants of context-specific target selection unknown","Mechanism integrating nuclear transcription and cytoplasmic Rho GTPase roles unclear"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0140110","term_label":"transcription regulator activity","supporting_discovery_ids":[3,6,11,21]},{"term_id":"GO:0003677","term_label":"DNA 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Key determinant of the keratinocyte proliferation-differentiation switch involved in appropriate epidermal development (By similarity). Plays a role in regulating mammary epithelial cell proliferation (By similarity). 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LPS stimulates the expression of LPS-binding protein in human oral keratinocytes in vitro.","date":"2012","source":"Innate immunity","url":"https://pubmed.ncbi.nlm.nih.gov/22736337","citation_count":30,"is_preprint":false},{"pmid":"12920575","id":"PMC_12920575","title":"Novel mutations in the IRF6 gene for Van der Woude syndrome.","date":"2003","source":"Human genetics","url":"https://pubmed.ncbi.nlm.nih.gov/12920575","citation_count":30,"is_preprint":false},{"pmid":"28357449","id":"PMC_28357449","title":"Myricetin protects cardiomyocytes from LPS-induced injury.","date":"2017","source":"Herz","url":"https://pubmed.ncbi.nlm.nih.gov/28357449","citation_count":30,"is_preprint":false},{"pmid":"7890395","id":"PMC_7890395","title":"Prolonged expression of lipopolysaccharide (LPS)-induced inflammatory genes in whole blood requires continual exposure to LPS.","date":"1995","source":"Infection and immunity","url":"https://pubmed.ncbi.nlm.nih.gov/7890395","citation_count":29,"is_preprint":false},{"pmid":"22002480","id":"PMC_22002480","title":"LPS-binding protein enables intestinal epithelial restitution despite LPS exposure.","date":"2012","source":"Journal of pediatric gastroenterology and nutrition","url":"https://pubmed.ncbi.nlm.nih.gov/22002480","citation_count":27,"is_preprint":false},{"pmid":"8524933","id":"PMC_8524933","title":"Lipopolysaccharide (LPS) binding protein catalyzes binding of LPS to lipoproteins.","date":"1995","source":"Progress in clinical and biological research","url":"https://pubmed.ncbi.nlm.nih.gov/8524933","citation_count":27,"is_preprint":false},{"pmid":"33234718","id":"PMC_33234718","title":"An Irf6-Esrp1/2 regulatory axis controls midface morphogenesis in vertebrates.","date":"2020","source":"Development (Cambridge, England)","url":"https://pubmed.ncbi.nlm.nih.gov/33234718","citation_count":25,"is_preprint":false},{"pmid":"9733616","id":"PMC_9733616","title":"Pulmonary LPS-binding protein (LBP) upregulation following LPS-mediated injury.","date":"1998","source":"The Journal of surgical research","url":"https://pubmed.ncbi.nlm.nih.gov/9733616","citation_count":25,"is_preprint":false},{"pmid":"28732181","id":"PMC_28732181","title":"IRF6 and SPRY4 Signaling Interact in Periderm Development.","date":"2017","source":"Journal of dental research","url":"https://pubmed.ncbi.nlm.nih.gov/28732181","citation_count":25,"is_preprint":false},{"pmid":"20472936","id":"PMC_20472936","title":"ABCA1 promotes the efflux of bacterial LPS from macrophages and accelerates recovery from LPS-induced tolerance.","date":"2010","source":"Journal of lipid research","url":"https://pubmed.ncbi.nlm.nih.gov/20472936","citation_count":25,"is_preprint":false},{"pmid":"38378697","id":"PMC_38378697","title":"Plasticity-induced repression of Irf6 underlies acquired resistance to cancer immunotherapy in pancreatic ductal adenocarcinoma.","date":"2024","source":"Nature communications","url":"https://pubmed.ncbi.nlm.nih.gov/38378697","citation_count":24,"is_preprint":false},{"pmid":"35983049","id":"PMC_35983049","title":"Sulforaphane diminishes moonlighting of pyruvate kinase M2 and interleukin 1β expression in M1 (LPS) macrophages.","date":"2022","source":"Frontiers in immunology","url":"https://pubmed.ncbi.nlm.nih.gov/35983049","citation_count":24,"is_preprint":false},{"pmid":"31943082","id":"PMC_31943082","title":"SPECC1L regulates palate development downstream of IRF6.","date":"2020","source":"Human molecular genetics","url":"https://pubmed.ncbi.nlm.nih.gov/31943082","citation_count":24,"is_preprint":false},{"pmid":"21045959","id":"PMC_21045959","title":"IRF6 Screening of Syndromic and a priori Non-Syndromic Cleft Lip and Palate Patients: Identification of a New Type of Minor VWS Sign.","date":"2010","source":"Molecular syndromology","url":"https://pubmed.ncbi.nlm.nih.gov/21045959","citation_count":24,"is_preprint":false},{"pmid":"30073169","id":"PMC_30073169","title":"LPS Induces mTORC1 and mTORC2 Activation During Monocyte Adhesion.","date":"2018","source":"Frontiers in molecular biosciences","url":"https://pubmed.ncbi.nlm.nih.gov/30073169","citation_count":23,"is_preprint":false},{"pmid":"12150986","id":"PMC_12150986","title":"Inhibition of LPS-induced activation of alveolar macrophages by high concentrations of LPS-binding protein.","date":"2002","source":"Biochemical and biophysical research communications","url":"https://pubmed.ncbi.nlm.nih.gov/12150986","citation_count":23,"is_preprint":false},{"pmid":"33362762","id":"PMC_33362762","title":"TLR4 Response to LPS Is Reinforced by Urokinase Receptor.","date":"2020","source":"Frontiers in immunology","url":"https://pubmed.ncbi.nlm.nih.gov/33362762","citation_count":22,"is_preprint":false},{"pmid":"28645193","id":"PMC_28645193","title":"IRF6 Regulates Alternative Activation by Suppressing PPARγ in Male Murine Macrophages.","date":"2017","source":"Endocrinology","url":"https://pubmed.ncbi.nlm.nih.gov/28645193","citation_count":22,"is_preprint":false},{"pmid":"31825181","id":"PMC_31825181","title":"PBX-WNT-P63-IRF6 pathway in nonsyndromic cleft lip and palate.","date":"2019","source":"Birth defects research","url":"https://pubmed.ncbi.nlm.nih.gov/31825181","citation_count":21,"is_preprint":false},{"pmid":"28767310","id":"PMC_28767310","title":"Disrupted IRF6-NME1/2 Complexes as a Cause of Cleft Lip/Palate.","date":"2017","source":"Journal of dental research","url":"https://pubmed.ncbi.nlm.nih.gov/28767310","citation_count":21,"is_preprint":false},{"pmid":"14563359","id":"PMC_14563359","title":"Distinct LPS-induced signals regulate LPS uptake and morphological changes in medfly hemocytes.","date":"2003","source":"Insect biochemistry and molecular biology","url":"https://pubmed.ncbi.nlm.nih.gov/14563359","citation_count":21,"is_preprint":false},{"pmid":"15013698","id":"PMC_15013698","title":"A novel mutation of the IRF6 gene in an Italian family with Van der Woude syndrome.","date":"2004","source":"Mutation research","url":"https://pubmed.ncbi.nlm.nih.gov/15013698","citation_count":21,"is_preprint":false},{"pmid":"23451037","id":"PMC_23451037","title":"Cell-autonomous and non-cell-autonomous roles for IRF6 during development of the tongue.","date":"2013","source":"PloS one","url":"https://pubmed.ncbi.nlm.nih.gov/23451037","citation_count":20,"is_preprint":false},{"pmid":"27014863","id":"PMC_27014863","title":"RIPK4 activates an IRF6-mediated proinflammatory cytokine response in keratinocytes.","date":"2016","source":"Cytokine","url":"https://pubmed.ncbi.nlm.nih.gov/27014863","citation_count":20,"is_preprint":false}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":51157,"output_tokens":5992,"usd":0.121676,"stage1_stop_reason":"end_turn"},"stage2":{"model":"claude-opus-4-8","input_tokens":14816,"output_tokens":5540,"usd":0.10629,"stage2_stop_reason":"end_turn"},"total_usd":0.227966,"stage1_batch_id":"msgbatch_01DUPACZi7AAWwuZiUjseQF4","stage2_batch_id":"msgbatch_018yt7chE3f48PEFtDwtj6AE","note":"batch pricing = 50% of standard"},"stage1_raw":"```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 2002,\n      \"finding\": \"Loss-of-function (nonsense/missense) mutations in IRF6 cause Van der Woude syndrome and popliteal pterygium syndrome. IRF6 mRNA is highly expressed along the medial edge of the fusing palate, tooth buds, hair follicles, genitalia and skin, establishing a direct role for IRF6 in orofacial and epithelial development.\",\n      \"method\": \"Mutation screening of VWS/PPS families; in situ expression analysis of Irf6 mRNA\",\n      \"journal\": \"Nature genetics\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — direct mutation identification in >45 families plus expression analyses; replicated across multiple independent cohorts in subsequent studies\",\n      \"pmids\": [\"12219090\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2006,\n      \"finding\": \"Mice homozygous for a missense mutation in Irf6 develop a hyperproliferative epidermis that fails to undergo terminal differentiation, resulting in soft tissue fusions. Compound heterozygosity for Irf6 and Sfn (14-3-3σ/stratifin) recapitulates similar keratinizing epithelial defects, placing Irf6 and Sfn in a shared genetic pathway controlling the keratinocyte proliferation-differentiation switch.\",\n      \"method\": \"Mouse genetics (homozygous missense knock-in); compound heterozygote epistasis analysis\",\n      \"journal\": \"Nature genetics\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — genetic epistasis with defined cellular phenotype (hyperproliferation/failure of terminal differentiation); compound heterozygote experiment provides independent mechanistic validation\",\n      \"pmids\": [\"17041603\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2009,\n      \"finding\": \"IRF6 is essential for oral epithelial differentiation and functions in a convergent molecular pathway with the Notch ligand Jagged2. IRF6 plays a key role in formation and maintenance of the oral periderm, with spatio-temporal regulation of this periderm being essential for appropriate palatal adhesion and fusion.\",\n      \"method\": \"Mouse genetic analysis (Irf6 mutant and Jagged2 mutant); histological and molecular characterization of palatal/oral epithelium\",\n      \"journal\": \"Human molecular genetics\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — genetic epistasis between Irf6 and Jagged2, with mechanistic characterization of periderm formation and palatal adhesion phenotype\",\n      \"pmids\": [\"19439425\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2011,\n      \"finding\": \"IRF6 is transcriptionally activated by p63 and, in turn, induces proteasome-mediated down-regulation of p63, forming a regulatory feedback loop that limits keratinocyte proliferative potential. ChIP-seq and gene expression profiling after siRNA-mediated IRF6 depletion revealed direct IRF6 target genes involved in cell cycle, differentiation, cell adhesion, and cell–cell contact. IRF6 down-regulation promotes invasive behavior in squamous cell carcinoma cells.\",\n      \"method\": \"ChIP-seq for IRF6 binding sites; siRNA knockdown; gene expression profiling; in vitro invasion assays; re-expression of IRF6 in SCC cells\",\n      \"journal\": \"Proceedings of the National Academy of Sciences of the United States of America\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — multiple orthogonal methods (ChIP-seq, siRNA KD, invasion assay, reintroduction rescue); single lab but mechanistically comprehensive\",\n      \"pmids\": [\"21807998\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2011,\n      \"finding\": \"IRF6 is induced during keratinocyte differentiation through a Notch-dependent mechanism and is a primary Notch target gene in keratinocytes and keratinocyte-derived SCC cells. Increased IRF6 expression contributes to Notch-driven regulation of growth/differentiation genes, while it is not required for induction of canonical Notch targets (p21WAF1/Cip1, Hes1, Hey1). Down-modulation of IRF6 counteracts differentiation and promotes ras-induced tumour formation.\",\n      \"method\": \"Notch gain-of-function; IRF6 siRNA knockdown; gene expression analysis; in vivo tumour formation assay\",\n      \"journal\": \"The EMBO journal\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — multiple orthogonal approaches (gain-of-function, knockdown, in vivo tumour model), distinct from but complementary to PMID 21807998\",\n      \"pmids\": [\"21909072\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2013,\n      \"finding\": \"TGFβ signaling regulates expression of Irf6, and IRF6 and SMAD4 synergistically regulate the fate of the medial edge epithelium (MEE) during palatal fusion. Haploinsufficiency of Irf6 in Smad4 conditional knockout mice causes compromised p21 expression and MEE persistence. Overexpression of Irf6 rescued p21 expression and MEE degeneration in Tgfbr2-deficient mice, establishing TGFβ-mediated Irf6 activity as responsible for MEE degeneration.\",\n      \"method\": \"Mouse genetics (conditional knockout, compound heterozygotes); Irf6 overexpression rescue experiment; p21 immunostaining\",\n      \"journal\": \"Development (Cambridge, England)\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — genetic epistasis with rescue experiment using Irf6 overexpression; multiple mutant combinations tested\",\n      \"pmids\": [\"23406900\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"RIPK4 kinase activity is required for mouse development; RIPK4 phosphorylates IRF6 at Ser413 and Ser424, priming IRF6 for activation. RNA-seq, ChIP-seq, and ATAC-seq of wild-type versus IRF6-deficient skin showed IRF6 is enriched at bivalent promoters and its deficiency causes defective expression of genes involved in lipid metabolism and tight junction formation, culminating in a severe epidermal barrier defect.\",\n      \"method\": \"Kinase-dead knock-in mouse; phosphosite mapping (Ser413/Ser424); RNA-seq; ChIP-seq; ATAC-seq; lipid composition analysis of stratum corneum\",\n      \"journal\": \"Nature\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 / Strong — phosphorylation site identified with mutagenesis context, combined with genome-wide chromatin and transcriptomic profiling and functional barrier assay; highly rigorous single study\",\n      \"pmids\": [\"31578523\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2015,\n      \"finding\": \"TLR3 stimulation of keratinocytes upregulates IRF6, which selectively promotes IL-23p19 expression while suppressing IFN-β expression. IRF6 silencing enhanced poly(IC)-inducible IFN-β mRNA and inhibited IL-23p19 mRNA. Co-transfection of IRF6 increased IL-23p19 promoter activity but inhibited IFN-β promoter activity, defining IRF6 as a specificity-determining downstream mediator of TLR3 signalling.\",\n      \"method\": \"siRNA silencing of IRF6; gene reporter (promoter) assays; co-immunoprecipitation; proximity ligation assay\",\n      \"journal\": \"Immunology and cell biology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — siRNA knockdown plus reporter assays in primary keratinocytes; single lab with two orthogonal approaches\",\n      \"pmids\": [\"26303210\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"IRF6 acts downstream of IRAK1 to stimulate expression of the pro-inflammatory cytokine IL-36γ in oral epithelial cells in response to Porphyromonas gingivalis (via TLR2). Gene silencing and promoter reporter experiments confirmed IRF6 drives IL-36γ transcription.\",\n      \"method\": \"Gene silencing (siRNA); promoter reporter assays; TLR2 agonist stimulation\",\n      \"journal\": \"Journal of immunology (Baltimore, Md. : 1950)\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — siRNA plus promoter assays; single lab, two orthogonal approaches\",\n      \"pmids\": [\"26819203\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"RIPK4 overexpression in oral keratinocytes induces CCL5 and CXCL11 expression in an IRF6-dependent manner. Gene silencing of RIPK4 or IRF6 abolished PKC pathway-induced CCL5/CXCL11 expression. Promoter reporter assays showed RIPK4 stimulates transactivation of CCL5 and CXCL11 promoters by IRF6, establishing a RIPK4→IRF6 signalling axis for proinflammatory cytokine induction in keratinocytes.\",\n      \"method\": \"RIPK4 overexpression; siRNA knockdown of RIPK4 and IRF6; gene reporter (promoter) assays; PKC pathway activation\",\n      \"journal\": \"Cytokine\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — overexpression, siRNA knockdown, and promoter reporter assays; single lab with multiple orthogonal methods\",\n      \"pmids\": [\"27014863\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2015,\n      \"finding\": \"IRF6 is a mediator of TGFβ3 during palatal fusion: ectopic IRF6 expression rescued shTgfβ3-induced fusion defects in palatal organ culture. IRF6 increases SNAI2 (Slug) expression and decreases epithelial markers (E-cadherin, Plakophilin, ZO-1), regulating epithelial–mesenchymal transition (EMT) during palatal fusion. Knockdown of Snai2 abolished the IRF6 rescuing effect.\",\n      \"method\": \"Palatal shelf organ culture; shRNA knockdown; ectopic IRF6 expression; RT-PCR and protein analysis of EMT markers\",\n      \"journal\": \"Scientific reports\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — organ culture rescue experiments with knockdown and overexpression; single lab, multiple approaches\",\n      \"pmids\": [\"26240017\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"IRF6 suppresses PPARγ transcription by binding IRF recognition sites upstream of the PPARγ coding region, thereby suppressing alternative (M2) macrophage polarization. ChIP confirmed IRF6 occupancy at the PPARγ locus. IRF6 knockdown enhanced M2 activation; IRF6 overexpression dramatically attenuated it.\",\n      \"method\": \"Gain- and loss-of-function (overexpression and siRNA); chromatin immunoprecipitation (ChIP); computational binding site prediction\",\n      \"journal\": \"Endocrinology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — ChIP plus gain/loss-of-function; single lab with orthogonal methods\",\n      \"pmids\": [\"28645193\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"A regulatory mutation (350dupA) in the IRF6 enhancer element MCS9.7 abrogates binding of p63 and E47 transcription factors to overlapping cis-motifs, significantly disrupting enhancer activity. The 350dupA simultaneously creates a CAAAGT motif bound by Lef1; Lef1/β-Catenin chimeric protein repressed MCS9.7-350dupA enhancer activity, demonstrating a dual loss-of-function/gain-of-function mechanism regulating IRF6 expression.\",\n      \"method\": \"Enhancer reporter assays in human cell cultures; transgenic mouse lacZ reporter assay; site-directed mutagenesis of binding motifs; Lef1 overexpression; electrophoretic mobility shift / binding assays\",\n      \"journal\": \"Human molecular genetics\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 / Strong — multiple orthogonal methods (cell reporter assay, transgenic mouse assay, mutagenesis, protein binding disruption) in a single rigorous study\",\n      \"pmids\": [\"24442519\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"IRF6 is identified as a direct protein interactor of NME1 and NME2 (regulators of Rho-type GTPases and E-cadherin endocytosis) by yeast two-hybrid screening and co-immunoprecipitation. The IRF6–NME interaction is enhanced by phosphorylation of key serine residues in the IRF6 C-terminus. CLP-associated IRF6 missense mutations disrupt NME binding and result in elevated Rac1 and RhoA activation. NME1 and NME2 co-localize with IRF6 in the cytoplasm of primary palatal epithelial cells in vivo.\",\n      \"method\": \"Yeast two-hybrid screen; co-immunoprecipitation; phosphorylation assays; Rac1/RhoA activation assays; co-localization in primary palatal epithelial cells\",\n      \"journal\": \"Journal of dental research\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — yeast 2-hybrid plus co-IP plus functional Rho GTPase readout; single lab with multiple orthogonal methods\",\n      \"pmids\": [\"28767310\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"Overexpression of Irf6 in mice causes exencephaly through suppression of Tfap2a and Grhl3 expression, while loss of Irf6 function causes curly tail with reduced Tfap2a and Grhl3 expression in tail tissues, demonstrating that a Tfap2a–Irf6–Grhl3 genetic pathway operates during neurulation as well as orofacial development.\",\n      \"method\": \"Transgenic Irf6 overexpression mouse model; Irf6 loss-of-function mouse; gene expression analysis of Tfap2a and Grhl3\",\n      \"journal\": \"Human molecular genetics\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — genetic gain- and loss-of-function with molecular readout of pathway targets; single lab\",\n      \"pmids\": [\"30689861\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"Loss of Irf6 in mice causes craniosynostosis and mandibular hypoplasia. Double heterozygous Irf6+/−;Twist1+/− embryos show severe mandibular hypoplasia with reduced EDN1, DLX5, DLX6 and HAND2 expression in mesenchymal cells, demonstrating an intercellular genetic interaction between Irf6 (expressed in epithelium) and Twist1 (expressed in mesenchyme). Exogenous EDN1 peptide partially rescued Meckel's cartilage abnormalities.\",\n      \"method\": \"Mouse genetics (single and double heterozygotes); gene expression analysis; mandibular explant culture with EDN1 rescue\",\n      \"journal\": \"Scientific reports\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — genetic epistasis with molecular readout and partial rescue; single lab\",\n      \"pmids\": [\"28769044\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2013,\n      \"finding\": \"Irf6 is expressed in Myf5+ (segmental paraxial mesoderm) cells of the developing tongue. Loss of Irf6 causes reduced and poorly organized Myf5+ cell lineage and aberrant cytoskeletal formation. Non-cell-autonomously, loss of Irf6 alters Bmp2, Bmp4, Shh, and Fgf10 signalling in the tongue, establishing both cell-autonomous (muscle differentiation/cytoskeletal organization) and non-cell-autonomous (cranial neural crest-derived inter-molar eminence) roles for IRF6.\",\n      \"method\": \"Lineage tracing (fate mapping); dual-label immunostaining; Irf6 knockout mouse analysis; molecular analysis of signalling pathway genes\",\n      \"journal\": \"PloS one\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — lineage tracing combined with KO analysis and molecular pathway readout; single lab\",\n      \"pmids\": [\"23451037\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"In zebrafish, irf6 regulates the expression of esrp1. Genetic disruption of irf6 and esrp1/2 in zebrafish results in cleft of the anterior neurocranium due to impaired chondrogenesis (not a neural crest migration defect). Mouse compound mutant analysis showed genetic interaction between Irf6 and Esrp1, with Irf6 heterozygosity reducing Esrp1/2 cleft severity, defining an Irf6–Esrp1/2 regulatory axis in midface morphogenesis.\",\n      \"method\": \"Zebrafish genetic knockouts; cranial neural crest lineage tracing; mouse compound mutant breeding; gene expression analysis\",\n      \"journal\": \"Development (Cambridge, England)\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — genetic epistasis across two species (zebrafish and mouse) with lineage tracing; single lab\",\n      \"pmids\": [\"33234718\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"SPECC1L expression is drastically reduced in Irf6 mutant palatal shelves, placing SPECC1L downstream of IRF6 in palatogenesis. Specc1l mutant mice display periderm layer abnormalities and transient oral epithelial adhesions similar to Irf6 hypomorphic mutants.\",\n      \"method\": \"Irf6 mutant mouse analysis; immunostaining for SPECC1L expression; phenotypic comparison of mutant palatal shelves\",\n      \"journal\": \"Human molecular genetics\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Weak — loss-of-function with molecular readout showing pathway placement; single lab, single method for the IRF6-SPECC1L relationship\",\n      \"pmids\": [\"31943082\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"In a mouse PDAC model, acquired resistance to immunotherapy is associated with EMT. EMT-transcription factors ZEB1 and SNAIL epigenetically and transcriptionally silence IRF6, rendering tumor cells resistant to the pro-apoptotic effects of TNF-α and insensitive to T-cell killing. This identifies IRF6 as a downstream mediator of EMT-driven immune evasion.\",\n      \"method\": \"Mouse PDAC tumor relapse model; EMT transcription factor manipulation; epigenetic analysis; TNF-α killing assays\",\n      \"journal\": \"Nature communications\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — in vivo tumor model plus mechanistic TNF-α sensitivity assay linking IRF6 silencing to apoptotic resistance; single lab\",\n      \"pmids\": [\"38378697\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2008,\n      \"finding\": \"IRF6 interacts with maspin (a known tumor suppressor) in mammary epithelial cells and functions to regulate cell cycle exit and entry into G0 quiescence, promoting mammary epithelial cell differentiation.\",\n      \"method\": \"Protein interaction (co-immunoprecipitation implied by 'interaction with maspin'); cell cycle analysis in mammary epithelial cells\",\n      \"journal\": \"Cell cycle (Georgetown, Tex.)\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — review/perspective piece describing prior interaction finding without detailed experimental reporting in this abstract; single method implied\",\n      \"pmids\": [\"18604160\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"IRF6 inhibits transcription of PKM2 and GLUT1 in glioma cells, thereby impairing glycolysis. SNHG14 lncRNA sequesters STAU1 to prevent IRF6 mRNA degradation via its 3' UTR, increasing IRF6 expression; Lin28A stabilizes SNHG14. Depleting Lin28A or SNHG14 or overexpressing IRF6 reduced xenograft tumor growth.\",\n      \"method\": \"siRNA/shRNA knockdown; overexpression; ChIP (IRF6 binding to PKM2/GLUT1 promoters); xenograft mouse model; RT-PCR and western blot\",\n      \"journal\": \"Cell death & disease\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — ChIP plus gain/loss-of-function plus in vivo xenograft; single lab with multiple orthogonal methods\",\n      \"pmids\": [\"32527996\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"IRF6 is a transcription factor that occupies a central node in epithelial differentiation: it is transcriptionally activated by p63 (via the MCS9.7 enhancer) and feeds back to suppress p63 via proteasomal degradation; it is phosphorylated at Ser413/Ser424 by the kinase RIPK4, which primes its transcriptional activity; it regulates a broad program of keratinocyte terminal differentiation (including lipid metabolism and tight junction genes), controls the proliferation-to-differentiation switch, operates downstream of TGFβ/SMAD4 and Notch signalling in palatal medial edge epithelium, interacts genetically with Jagged2, Sfn, Esrp1/2, Twist1, and GRHL3 during craniofacial and neural tube morphogenesis, drives IL-36γ and CCL5/CXCL11 expression downstream of TLR2/3 and RIPK4 in keratinocytes, suppresses PPARγ to limit M2 macrophage polarization, and is epigenetically silenced by EMT transcription factors to render tumor cells resistant to TNF-α-mediated killing.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"IRF6 is a transcription factor that occupies a central node in epithelial differentiation and craniofacial morphogenesis, where it governs the keratinocyte proliferation-to-differentiation switch and the loss-of-function mutations cause the Mendelian orofacial clefting disorders Van der Woude syndrome and popliteal pterygium syndrome [#0]. IRF6 sits within a regulatory circuit in which it is transcriptionally activated by p63 through the MCS9.7 enhancer and reciprocally drives proteasomal down-regulation of p63, a feedback loop that limits keratinocyte proliferative potential and restrains invasive behavior [#3, #12]; it is also a primary Notch target gene whose induction promotes terminal differentiation and opposes ras-driven tumorigenesis [#4]. Full transcriptional activation requires priming by the kinase RIPK4, which phosphorylates IRF6 at Ser413 and Ser424; through this axis IRF6 occupies bivalent promoters and directs a terminal-differentiation program including lipid metabolism and tight junction genes necessary for the epidermal barrier [#6]. During palatal fusion IRF6 acts downstream of TGFβ/SMAD4 signalling to drive p21 expression and medial edge epithelium degeneration [#5], operates in convergent pathways with the Notch ligand Jagged2 and the stratifin Sfn to maintain the oral periderm [#1, #2], and participates in genetic networks with Twist1, Esrp1/2, Tfap2a–Grhl3, and SPECC1L that pattern craniofacial and neural tube development [#14, #15, #17, #18]. Beyond development, IRF6 functions as a specificity-determining mediator of innate immune signalling in keratinocytes, driving IL-36γ, CCL5/CXCL11, and IL-23p19 expression downstream of TLR2/TLR3 and RIPK4 [#7, #8, #9], and it suppresses PPARγ to constrain M2 macrophage polarization [#11]. In cancer, epigenetic silencing of IRF6 by the EMT factors ZEB1 and SNAIL confers resistance to TNF-α-mediated apoptosis and T-cell killing [#19].\",\n  \"teleology\": [\n    {\n      \"year\": 2002,\n      \"claim\": \"Establishing that IRF6 mutations cause human orofacial clefting syndromes defined IRF6 as an essential regulator of epithelial and craniofacial development, anchoring all subsequent mechanistic work.\",\n      \"evidence\": \"Mutation screening of Van der Woude/popliteal pterygium syndrome families plus in situ expression of Irf6 mRNA\",\n      \"pmids\": [\"12219090\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Molecular target genes of IRF6 not yet defined\", \"Upstream regulators of IRF6 expression unknown\"]\n    },\n    {\n      \"year\": 2006,\n      \"claim\": \"Mouse genetics showed that IRF6 controls the keratinocyte proliferation-to-differentiation switch, explaining the epithelial fusions seen in disease.\",\n      \"evidence\": \"Homozygous missense knock-in mouse and Irf6;Sfn compound heterozygote epistasis with hyperproliferation/differentiation-failure phenotype\",\n      \"pmids\": [\"17041603\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Molecular mechanism by which IRF6 enforces differentiation not resolved\", \"How Sfn and IRF6 intersect biochemically unknown\"]\n    },\n    {\n      \"year\": 2009,\n      \"claim\": \"IRF6 was placed in a convergent pathway with the Notch ligand Jagged2 controlling oral periderm formation, linking it to palatal adhesion and fusion.\",\n      \"evidence\": \"Mouse genetic analysis of Irf6 and Jagged2 mutants with histological characterization of periderm\",\n      \"pmids\": [\"19439425\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Direct biochemical link between IRF6 and Notch signalling not established here\", \"Periderm-specific IRF6 targets unidentified\"]\n    },\n    {\n      \"year\": 2011,\n      \"claim\": \"Defining the p63→IRF6→p63 feedback loop and genome-wide IRF6 targets converted IRF6 from a genetic locus into a characterized transcription factor restraining proliferation and invasion.\",\n      \"evidence\": \"ChIP-seq, siRNA knockdown, gene expression profiling, invasion assays and IRF6 re-expression in SCC cells; plus Notch gain/loss-of-function and in vivo tumour assays\",\n      \"pmids\": [\"21807998\", \"21909072\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Mechanism of IRF6-driven p63 proteasomal degradation not detailed\", \"Post-translational control of IRF6 activity not yet known\"]\n    },\n    {\n      \"year\": 2013,\n      \"claim\": \"Genetic and rescue experiments established TGFβ/SMAD4 as an upstream activator of IRF6 driving p21 expression and medial edge epithelium degeneration during palatal fusion.\",\n      \"evidence\": \"Mouse conditional knockouts, compound heterozygotes, and Irf6 overexpression rescue of Tgfbr2-deficient MEE persistence with p21 immunostaining\",\n      \"pmids\": [\"23406900\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Direct transcriptional regulation of Irf6 by SMAD4 not demonstrated\", \"Whether p21 is a direct IRF6 target unresolved\"]\n    },\n    {\n      \"year\": 2013,\n      \"claim\": \"Lineage tracing revealed both cell-autonomous (muscle/cytoskeletal) and non-cell-autonomous roles for IRF6 in tongue development, broadening its developmental scope beyond epithelium.\",\n      \"evidence\": \"Fate mapping, Irf6 knockout analysis, and molecular readout of Bmp/Shh/Fgf signalling in tongue\",\n      \"pmids\": [\"23451037\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Mechanism of non-cell-autonomous signalling effects unknown\", \"Single lab, correlative pathway readouts\"]\n    },\n    {\n      \"year\": 2014,\n      \"claim\": \"Dissection of the MCS9.7 enhancer showed how a regulatory mutation reprograms IRF6 expression through dual loss and gain of transcription-factor binding, explaining noncoding clefting risk.\",\n      \"evidence\": \"Enhancer reporter assays, transgenic mouse lacZ assay, site-directed mutagenesis, and binding assays for p63/E47/Lef1\",\n      \"pmids\": [\"24442519\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"In vivo contribution of Lef1/β-Catenin repression not quantified\", \"Other enhancers controlling IRF6 not mapped\"]\n    },\n    {\n      \"year\": 2015,\n      \"claim\": \"Functional studies positioned IRF6 as a downstream effector of TGFβ3-driven EMT during palatal fusion, acting through SNAI2.\",\n      \"evidence\": \"Palatal organ culture rescue, shRNA knockdown, ectopic IRF6 expression, and EMT marker analysis\",\n      \"pmids\": [\"26240017\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Whether SNAI2 is a direct IRF6 target not shown\", \"Reconciliation with IRF6's anti-EMT role in tumors unresolved\"]\n    },\n    {\n      \"year\": 2015,\n      \"claim\": \"IRF6 was identified as a specificity-determining switch in TLR3 signalling, biasing keratinocytes toward IL-23p19 and away from IFN-β.\",\n      \"evidence\": \"siRNA silencing, promoter reporter assays, co-IP and proximity ligation in primary keratinocytes\",\n      \"pmids\": [\"26303210\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"IRF6 binding partner mediating promoter selectivity not defined\", \"In vivo relevance to skin immunity not tested\"]\n    },\n    {\n      \"year\": 2016,\n      \"claim\": \"Two studies extended IRF6 into innate immune cytokine induction, defining IRAK1→IRF6 (IL-36γ) and RIPK4→IRF6 (CCL5/CXCL11) axes in epithelial inflammation.\",\n      \"evidence\": \"siRNA silencing and promoter reporter assays with TLR2/PKC stimulation and RIPK4 overexpression in oral keratinocytes\",\n      \"pmids\": [\"26819203\", \"27014863\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Direct IRF6 binding to cytokine promoters not all confirmed by ChIP\", \"Single lab reporter-based evidence\"]\n    },\n    {\n      \"year\": 2017,\n      \"claim\": \"Identification of NME1/NME2 as direct IRF6 interactors revealed a cytoplasmic, phosphorylation-dependent function linking IRF6 to Rho GTPase regulation, beyond its nuclear transcriptional role.\",\n      \"evidence\": \"Yeast two-hybrid, co-IP, phosphorylation and Rac1/RhoA activation assays, and co-localization in primary palatal epithelial cells\",\n      \"pmids\": [\"28767310\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Functional consequence of cytoplasmic IRF6 in vivo unclear\", \"How nuclear vs cytoplasmic IRF6 pools are partitioned unknown\"]\n    },\n    {\n      \"year\": 2017,\n      \"claim\": \"Epistasis with Twist1 demonstrated an epithelial-to-mesenchymal intercellular signalling role for IRF6 acting through EDN1/DLX/HAND2 in craniofacial skeletal development.\",\n      \"evidence\": \"Mouse single/double heterozygotes, gene expression analysis, and EDN1 rescue of mandibular explants\",\n      \"pmids\": [\"28769044\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Secreted mediator transmitting the epithelial-mesenchymal signal not identified\", \"Direct vs indirect regulation of EDN1 unknown\"]\n    },\n    {\n      \"year\": 2017,\n      \"claim\": \"IRF6 was shown to directly repress PPARγ transcription, constraining M2 macrophage polarization and extending IRF6 function to immune cell programming.\",\n      \"evidence\": \"Gain/loss-of-function and ChIP confirming IRF6 occupancy at the PPARγ locus\",\n      \"pmids\": [\"28645193\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Physiological context of macrophage IRF6 expression unclear\", \"Single lab\"]\n    },\n    {\n      \"year\": 2019,\n      \"claim\": \"Identification of RIPK4 as the kinase phosphorylating IRF6 at Ser413/Ser424 provided the priming mechanism for IRF6 activation and tied IRF6 directly to bivalent-promoter control of the epidermal barrier program.\",\n      \"evidence\": \"Kinase-dead knock-in mouse, phosphosite mapping, RNA-seq/ChIP-seq/ATAC-seq, and stratum corneum lipid/barrier assays\",\n      \"pmids\": [\"31578523\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Structural basis of phospho-priming not resolved\", \"How phosphorylation alters IRF6 DNA binding/coactivator recruitment unknown\"]\n    },\n    {\n      \"year\": 2019,\n      \"claim\": \"Gain- and loss-of-function established a Tfap2a–Irf6–Grhl3 genetic pathway operating in neurulation, generalizing IRF6's morphogenetic role to neural tube closure.\",\n      \"evidence\": \"Transgenic Irf6 overexpression and loss-of-function mice with Tfap2a/Grhl3 expression analysis\",\n      \"pmids\": [\"30689861\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Direct transcriptional regulation among pathway members not demonstrated\", \"Single lab\"]\n    },\n    {\n      \"year\": 2020,\n      \"claim\": \"Cross-species genetics defined an Irf6–Esrp1/2 axis and placed SPECC1L downstream of IRF6 in midface and periderm morphogenesis, refining the developmental network.\",\n      \"evidence\": \"Zebrafish knockouts with neural crest lineage tracing, mouse compound mutants, and Irf6-mutant SPECC1L immunostaining\",\n      \"pmids\": [\"33234718\", \"31943082\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Direct vs indirect regulation of esrp1/SPECC1L by IRF6 not established\", \"Single method for the IRF6-SPECC1L link\"]\n    },\n    {\n      \"year\": 2020,\n      \"claim\": \"IRF6 was shown to repress glycolytic genes PKM2 and GLUT1 in glioma and to be controlled post-transcriptionally by a Lin28A/SNHG14/STAU1 axis, extending IRF6 to tumor metabolism.\",\n      \"evidence\": \"ChIP, knockdown/overexpression, and xenograft assays\",\n      \"pmids\": [\"32527996\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Relevance of glioma metabolic role to epithelial IRF6 biology unclear\", \"Single lab\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"IRF6 was identified as a downstream mediator of EMT-driven immune evasion, with ZEB1/SNAIL silencing of IRF6 conferring resistance to TNF-α apoptosis and T-cell killing.\",\n      \"evidence\": \"Mouse PDAC relapse model, EMT-TF manipulation, epigenetic analysis, and TNF-α killing assays\",\n      \"pmids\": [\"38378697\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"IRF6 target genes mediating TNF-α sensitivity not defined\", \"Whether human tumors recapitulate this silencing not shown\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"The structural and biochemical basis by which phosphorylation, cytoplasmic interactors, and chromatin context switch IRF6 between activator and repressor functions across epithelial, immune, and tumor contexts remains unresolved.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No structural model of activated IRF6 on target promoters\", \"Determinants of context-specific target selection unknown\", \"Mechanism integrating nuclear transcription and cytoplasmic Rho GTPase roles unclear\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0140110\", \"supporting_discovery_ids\": [3, 6, 11, 21]},\n      {\"term_id\": \"GO:0003677\", \"supporting_discovery_ids\": [3, 11, 12]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005634\", \"supporting_discovery_ids\": [3, 6]},\n      {\"term_id\": \"GO:0005829\", \"supporting_discovery_ids\": [13]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-1266738\", \"supporting_discovery_ids\": [0, 2, 5, 14, 17]},\n      {\"term_id\": \"R-HSA-74160\", \"supporting_discovery_ids\": [3, 6, 11]},\n      {\"term_id\": \"R-HSA-168256\", \"supporting_discovery_ids\": [7, 8, 9, 11]},\n      {\"term_id\": \"R-HSA-1640170\", \"supporting_discovery_ids\": [1, 3, 20]}\n    ],\n    \"complexes\": [],\n    \"partners\": [\"TP63\", \"RIPK4\", \"SMAD4\", \"NME1\", \"NME2\", \"SFN\"],\n    \"other_free_text\": []\n  }\n}","audit_flag":null,"evaluation":{"pairwise":"win","faith_supported":6,"faith_total":6,"faith_pct":100.0}}