Figure 1.

Transient KD of YAP impairs differentiation of ESCs.A, α1T–GFP stable cell line transfected with Stealth siRNA to silence YAP expression (KD YAP) showed a decrease in neuroectodermal differentiation efficacy with respect to KD CTR cells. Representative images are shown. The scale bar represents 100 μm. B, representative immunostaining of neural marker β3-tubulin (red) in α1T–GFP (green) at final stage (T14) of differentiation showing a dramatic reduction in postmitotic neuronal differentiation. The scale bar represents 100 μm. C, quantitative PCR analysis of neuronal marker gene expression upon differentiation of YAP KD cells. Data are shown as fold changes with respect to KD CTR cells. ∗p < 0.05, ∗∗p < 0.01. D, YAP expression profile over neuroectodermal differentiation by Western blot or E, quantitative PCR. F, YAP KD in mlc2v–GFP stable cell line showed a strong reduction of mesodermal differentiation efficiency compared with KD CTR cells. Representative immunostainings are shown. The scale bar represents 100 μm. G, percentage of beating areas at final stage of differentiation (T8), demonstrating a significant (∗∗∗p < 0.01) decrease in generation of mature cardiomyocytes upon YAP KD. For each data set, averaged numbers from biological triplicates were used for statistics. Error bars indicate mean ± SEM. CTR, control; KD, knockdown; YAP, Yes-associated protein.