Fig 2: TEAD interacts with ERa and promotes ERa signaling activity.a High levels of TEAD4 correlate with poor survival in endocrine therapy breast cancer patients based on https://kmplot.com/analysis/. b The effect of TEAD4 RNAi on TEAD4 and ERa expression in MCF-7. c, d TEAD4 RNAi inhibited ER+ breast cancer cell growth. MCF-7 (c) and T47D (d) cells transfected with control or TEAD4 siRNA were analyzed using the WST-1 assay at indicated time points. e TEAD4 depletion reduced ERa target gene expression. MCF-7 cells transfected with TEAD4 siRNA were grown for 48 h, followed by RT-qPCR analysis of the indicated ERa target genes. f ChIP-qPCR assay showed that TEAD4 RNAi decreased ERa recruitment to its target gene promoters/enhancers. g Western blot analysis of ERa and TEAD4 expression in control or MCF-7 cells stably expressing RNAi-insensitive TEAD4 (TEAD4INS) and transfected with control or TEAD4 siRNA. h TEAD4INS restored the expression of ERa target genes in TEAD4 depleted MCF-7 cells. i ChIP-qPCR assay showed that TEAD4 and ERa co-occupied on ERa target promoters/enhancers in MCF-7 cells. j Co-IP experiments revealed that TEAD4 formed a complex with ERa in MCF-7 cells. k, l TEAD4 (k) or ERa (l) domain structure and deletion mutants used for Co-IP experiments. m–o TEAD4 interacted with the AF2 domain of ERa through its YAP-binding domain. HEK293T cells were transfected with the indicated TEAD4 and ERa constructs, followed by Co-IP and western blot analyses. p ERa recruitment to its target gene promoters/enhancers is enhanced by TEAD41-434 but not by TEAD4 deletion mutants TEAD41-220 or TEAD4131-434. HEK293T cells were transfected with HA-ERa together and the indicated TEAD4 constructs, followed by ChIP-qPCR analysis of HA-ERa binding to ERa target promoters/enhancers. q, r AF2 is required for TEAD4 to facilitate ERa binding to its target promoters/enhancers. HEK293T cells were transfected with GFP-TEAD4 and the indicated ERa constructs, followed by ChIP-qPCR analysis of occupancy of ERa or its deletion mutants on ERa target promoters/enhancers. Two-sided log-rank test were used for a. Results shown in b–j, m–r are representative of three independent experiments. Data are means ± s.d. Two-sided, unpaired t-test for c–f, h, i, p–r. Source data are provided in the Source Data file.

Fig 3: YAP inhibits ERa-TEAD interaction and hormone-independent ESR1 expression.a Coexpression of YAP with ERa and TEAD4 disrupted their association. HEK293T cells were transfected with fixed amounts of GFP-TEAD4 and HA- ERa and increasing amounts of Flag-YAP, followed by Co-IP and western blot analyses. b, c YAP inhibited TEAD4-mediated enhancement of ERa binding to its target promoters/enhancers. HEK293T cells were transfected with the indicated constructs, followed by western blot analysis (b) or ChIP-qPCR analysis for HA-ERa binding to its target promoters/enhancers (c). d XUM-MP-1 dissociated ERa from TEAD4 while promoted YAP/TEAD4 association. MCF-7 cells were treated with 2 µM XMU-MP-1 for the indicated time, followed by Co-IP and western blot analyses. e YAP inhibited ERa-TEAD4 association by binding to TEAD4. Control or MCF-7 cells expressing Tet-O-YAP5SA or Tet-O-YAP5SAS94A were treated with 0.2 µg/ml Dox for 10 h, followed by Co-IP and western blot analyses. f–h XUM-MP-1 increased the promoter/enhancer occupancy of YAP to ERa target genes including ESR1. MCF-7 cells were treated with 3 µM XMU-MP-1 for 6 h, followed by ChIP-qPCR analysis for ERa (f), TEAD4 (g), YAP (h) binding to ERa target promoters/enhancers. i Ectopic expression of active form of YAP increased YAP binding to ERa target promoters/enhancers. MCF-7 cells expressing Tet-O-YAP5SA were treated with 0.2 µg/ml Dox for the indicated time, followed by ChIP-qPCR analysis for YAP binding to ERa target promoters/enhancers. j, k Aggregate plots showing the normalized tag density of ERa (j) and YAP1 (k) ChIP-seq data for the ERa/TEAD sites in MCF-7 cells treated with vehicle or XMU-MP-1. l XUM-MP-1 inhibited hormone-independent transcription of ESR1 and ERa target genes. MCF-7 cells grown in hormone-depleted medium were treated 3 µM XMU-MP-1 for 6 h, followed by RT-qPCR analysis of the indicated genes. m YAP5SA but not YAP5SAS94A inhibited hormone-independent transcription of ESR1 and other ERa target genes. Control or MCF-7 cells expressing Tet-O-YAP5SA or Tet-O-YAP5SAS94A were treated with 0.2 µg/ml Dox for 16 h, followed by RT-qPCR analysis of the indicated genes. Results shown in a–i and l, m are representative of three independent experiments. Data are means ± s.d. Two-sided, unpaired t-test for c, f–i, l, m. Source data are provided in the Source Data file.