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. Author manuscript; available in PMC: 2018 Jan 28.
Published in final edited form as: Nat Cell Biol. 2017 Jul 28;19(8):996–1002. doi: 10.1038/ncb3581

Figure 4. TEAD inhibition restricts YAP-driven cancer cell growth.

Figure 4

a, b, Effect of osmotic stress on TEAD and YAP cytoplasmic sequestration in MSTO-211H mesothelioma cells. Note that unlike HEK293A (Fig. 1a), serum starvation, glucose starvation, and latruculin B did not induce YAP cytoplasmic localization because of Lats2 mutation in MSTO-211H cells. Only NaCl treatment elicited TEAD and YAP cytoplasmic translocation as detected by immunofluorescence (a) despite constitutive YAP dephosphorylation as detected by Western blot (b). c, Stress-induced TEAD inhibition suppresses anchorage independent growth. Osmotic stress inhibited colony formation in control, but not TEAD1/4-VP16 expressing MSTO-211H cells. d, e, Immunoprecipitation shows p38 cannot bind TEAD1-VP16 (d) or TEAD4-VP16 (e). f, Western blotting of YAP phosphorylation status in UM cell lines 92.1 and OCM1. g, Immunostaining of TEAD and YAP/TAZ in UM cell lines 92.1 and OCM1 upon osmotic stress. Note that YAP displays cytoplasmic staining under normal condition in OCM1 cells. Scale bars in a and g are 20μm. h, Differential effect of TEAD inhibition on anchorage-independent growth of GNAQ-mutant and BRAF-mutant UM cells. Stress-induced TEAD inhibition ablated colony formation in all GNAQ-mutant cell lines, whereas BRAF-mutant cell growth was insensitive. i, Western blot for PARP cleavage in 92.1 and OCM1 cells. NaCl stimulation induces apoptosis in 92.1 but not OCM1. Pretreatment with p38 inhibitor rescues cells from osmotic stress-induced apoptosis. j, TEAD1/4 -VP16 rescues p38-induced inhibition of colony formation of MSTO-211H cells. k, Quantification of (j). n=3 biological replicates. Data are presented as mean ± s.e.m. *p < 0.05; **p < 0.01; p values were determined using one-way ANOVA followed by Tukey’s multiple comparison test. l, TEAD1/4-VP16 rescues p38-induced inhibition of MSTO-211H in vivo tumor xenograft growth. Nude mice were injected with control, p38, or p38 + TEAD1/4-VP16 expressing MSTO-211H cells and tumor growth was measured at the indicated times. Data are presented as mean ± s.e.m. n = 4 mice per group. **p < 0.01; ***p < 0.001; p values were determined using two-way ANOVA. m, Nude mice were injected with control, p38, or p38 + TEAD1/4-VP16 expressing MSTO-211H cells and tumors were harvested after 4 weeks. Only three tumors developed in the p38 group. Scale bar, 10mm.