Figure 6

CNN3 deficiency induces activation of the Yap/Taz and fibrogenic responses in lens epithelial cell cultures. siRNA-mediated downregulation of CNN3 expression in mouse lens epithelial cells as described in the Methods section results in significant decreases (*P < 0.01; Student t test) in the levels of phospho-Yap and total Yap, together with significant increase (*P < 0.01; Student t test) in the protein levels of Taz compared to controls cells treated with scrambled siRNA, based on immunoblotting analysis (panel A) and densitometric quantification (panel B). There was robust nuclear translocation of Yap in CNN3 deficient lens epithelial cells based on immunofluorescence analysis (panel C). Significant increases (*P < 0.05; Student t test) were also observed in the levels of CTGF and fibronectin (immunoblotting, panel D and densitometry, panel E) and fibronectin production (immunofluorescence; panel F) in CNN3 deficient lens epithelial cells compared to respective control cells. Scale bars indicate image magnification and GAPDH was immunoblotted for loading control. Sc and Scr siRNA: Scrambled siRNA.