Skip to main content
. 2019 Jul 9;9(17):4993–5008. doi: 10.7150/thno.35665

Figure 2.

Figure 2

CD44 acts through RhoA modulating YAP activation induced by matrix stiffness stimuli. (A-D) NIH-3T3 fibroblasts were transfected with NC or CD44 siRNA on stiff (60 kappa) gel-coated coverslips for 48 hours. (A) NIH-3T3 fibroblasts were immunostained with an antibody recognizing YAP. The percentage of cells with predominantly nuclear YAP staining was quantified. Nuclei were counter-stained with DAPI (n=3; ***, P < 0.001). Scale bar, 50 μm. (B-D) Western blot analysis of YAP expression and Rho family proteins (RhoA, Rac and Cdc42). β-Actin was used as a loading control. Quantification of YAP (C) and Rho family proteins (D) is shown. (E-F) NIH-3T3 fibroblasts were transfected with NC or RhoA, Rac1, or Cdc42 siRNAs on stiff (60 kappa) gel-coated coverslips for 48 hours, immunostained with phalloidine to visualize F-actin (E), and an antibody recognizing YAP to visualize YAP localization (F) (n=3; ***, P < 0.001). The percentage of cells with predominantly nuclear YAP staining was quantified. Scale bar, 50 μm. (G-I) NIH-3T3 fibroblasts were transfected with NC or RhoA (G), Rac1 (H), and Cdc42 (I) siRNAs. Cell lysates were subjected to immunoblotting with YAP antibody. β-Actin was used as a loading control. Quantification of YAP level is shown. (C-D and G-I) Data shown are representative of three independent experiments. Error bars indicate mean ± SD (**, P < 0.01; ***, P < 0.001; NS, not significant).