Figure 3

Changes in cellular tension lead to YAP activation. (A,C) Images of Blebbistatin or ML-7 treated mESCs in FN–coated micro-wells (magenta) with 15 and 35 µm base area edge length, respectively. Actin staining (green) demonstrates a large spreading area. Insets show 3D reconstruction of the nuclei of inhibitor treated cells in small micro-wells. Scale bars: 10 µm. (B,D) CDR of inhibitor treated mESCs does not differ between small and large micro-wells. The no-treatment control is shown in Fig. 1F. (E) Images of mESCs in small and large micro-wells four hours after seeding. The outline of the cell was determined by the actin signal (yellow line). The nuclear outline was determined by DAPI staining (blue line). Scale bars: 10 µm. (F) Quantification of the nuclear to cytoplasmic (n/c) YAP ratio in control cells. (G and H) Quantification of the n/c YAP ratio in inhibitor treated mESCs. Graphs show mean ± one s.d. from N = 5, 4, 3, respectively, independent experiments. The number of analyzed cells is given in the individual bars; p value was determined by two-tailed student’s t-test.