Extended Data Fig. 5: YAP/TAZ silencing sensitizes tumor cells to heat shock.

a, Verification of YAP/TAZ siRNA knockdown in B16-OVA cells by Western blot and RT-PCR. Presented are mean ± s.d., n = 3 biologically independent samples. Two-way ANOVA test. b, YAP/TAZ knockdown reduces B16-OVA cell viability in response to heat shock. Cells were transfected with control or two independent YAP and TAZ siRNAs, subjected to heat shock at 45 °C or 46 °C for 1 h, then recovered at 37 °C for 24 h. Cell viability was determined by CCK8 assays. Data are mean ± s.d.; n = 3 biologically independent experiments. Two-way ANOVA test. ns, not significant; NT, no treatment. c, YAP/TAZ knockdown promotes B16-OVA apoptosis. Images are representative of two independent experiments with similar results. d, More cell debris and apoptotic bodies in YAP/TAZ knockdown cells. B16-OVA cells transfected with siRNA, heat shocked at 45^oC for 1 h, and recovered at 37^oC for 6 h. Cells were subjected to microscopic examination for morphology. Representative pictures from three independent samples are shown. Scale bar, 100 μm. e, YAP/TAZ knockdown compromises HSP induction by heat shock. Cells were similarly treated as in (d). mRNAs were quantified by RT-PCR. Data are mean ± s.d.; n = 3 biologically independent samples. Two-way ANOVA test. ns, not significant. Only siRNA#2 was chosen in this experiment. f, g, YAP/TAZ knockdown reduces SCC7 cell viability upon heat shock. CCK8 assay (f) and Annexin V staining (g). Data presented as mean ± s.d.; n = 3 biologically independent experiments. One-way ANOVA test. h, Verification of YAP/TAZ siRNA knockdown in HCT116 tumor cells. Immunoblotting has been performed two times with similar results. i, YAP/TAZ knockdown HCT116 cells are more sensitive to heat shock stress. CCK8 assay was performed to measure cell viability. Data are presented as mean ± s.d.; n = 3 biologically independent experiments. One -way ANOVA test. Source data are available online.

Source data