Fig. 1
YAP translocate to the nucleus in trophoblast stem cells. A Western blot analysis of YAP and pYAPser127 using cell lysates prepared from TSCs (stem cells) and differentiated cells. GAPDH was used as an endogenous control. B Densitometric analysis of protein bands from A using NIH ImageJ software. C Western blot analysis of YAP in cytoplasmic and nuclear fraction of TSCs (stem cells) and differentiated cell lysate. GAPDH and Histone H3 were used as the cytoplasmic and membrane marker, respectively. D Densitometric quantification of YAP bands from C showing the relative amount of YAP in the nucleus and cytoplasm of TSCs and differentiated trophoblast cells. Normalization of nuclear and cytoplasmic YAP was done using Histone H3 and GAPDH, respectively. E Confocal photomicrographs of YAP (red) immunostained in TSCs and trophoblast cells on the 6th day of differentiation. Boxed areas on the right panel show extensive localization of YAP in the nucleus of TSCs and cytoplasm of differentiated cells. The nuclei were counterstained using Hoechst (blue). Scale bar: 20 µm. Magnification: × 60. F Ratio of the mean fluorescence intensity in the nucleus relative to the cytoplasm of TSCs and differentiated cells from E is represented as Corrected Total Cell Fluorescence (CTCF). Values are represented as mean ± SEM from three independent biological replicates and n = 10 in each replicate. **p < 0.005; ***p < 0.0005