Figure 6. WNT5A/B Are YAP/TAZ-TEAD-Mediated Wnt Target Genes.

(A) TEAD induce Wnt5a/b and inhibit Wnt/β-catenin signaling. Stable MCF10A-4SA and 4SA/S51A cells were serum-starved for 16 hr then stimulated with Wnt3a (100 ng/ml).

(B) WNT5A promoter region contains two TEAD binding sequences (TBS; GGAATG). Arrows underline primers used to amplify the region with each TBS.

(C) TEAD1 binds WNT5A promoter. ChIP experiment using anti-TEAD1 antibody was performed in TAZ-4SA-MCF10A cells. CTGF, positive control; GAPDH and WNT5A–CR (coding region), negative controls. Data are presented as mean ± SEM. ^*IgG versus anti-TEAD1 antibody, P < 0.05 (Student’s t-test).

(D) Activation of YAP/TAZ-Wnt5a/b axis by FZD1/ROR1. Stable MCF10A–FZD1/ROR1 cells were stimulated with Wnt3a (100 ng/ml). Note that Wnt3a induces Wnt5a/b accumulation in control cells (lanes 1–3).

(E) VP suppresses FZD1/ROR1-induced WNT5A gene expression. TEAD target genes, WNT5A and CTGF, and TCF target gene MYC mRNA were measured in MCF10A–FZD1/ROR1 (F+R) cells after VP (7 mM) treatment for 0, 2, 4 hr. Data are presented as mean ± SEM. *Compared to no VP treatment, P < 0.05 (Student’s t-test).

(F and G) TEAD mediates Wnt3a–induced WNT5A and WNT5B gene expression. Wnt3a (100 ng/ml) treatment for 0, 2, 4 hr induced mRNA expression of WNT5A (F) and WNT5B (G) in MCF10A and 3T3-L1 cells, respectively. In TEAD knockdown cells, Wnt3a–induced WNT5A/B expression was impaired. Data are presented as mean ± SEM. *Control versus shTEAD at 4 hr, P < 0.05 (Student’s t-test).

(H) Effect of oncogenic PIK3CA mutations on YAP/TAZ activation. PIK3CA-E545K and H1047R knockin MCF10A were starved for 16 hr then harvested.

(I) VP (7 µM) suppresses PIK3CA-H1047R–induced WNT5A and CTGF gene expression. Data are presented as mean ± SEM. *Compared to no VP treatment, P < 0.05 (Student’s t-test).

(J) VP (7 µM) induces β-catenin/TCF target genes in PIK3CA-H1047R knockin cells.

See also Figure S6.