Figure 5.

FAF1 alters the integrity of adherens junction. (A) HeLa cells were transfected with control siRNA or FAF1 siRNA #2 and replated on coverslips for 72 h. Cells were fixed and stained with β-catenin to investigate the integrity of adherens junction. Scale bar, 10 μm. (B) The fluorescence intensities of β-catenin are represented along the dashed line in A. Normalization was performed with respect to both end points of the dashed line in three different experiments. (C) HeLa cells were transfected with control siRNA or FAF1 siRNA for 48 h and replated on coverslips in a low-calcium medium. After 30 h, cells were returned to the normal-calcium medium to initiate junction assembly. The cells were then fixed and stained with β-catenin at the indicated time points. Scale bar, 10 μm. (D) HeLa cells were transfected with control siRNA or FAF1 siRNA for 72 h and lysed with gel sample buffer, and the lysates were analysed by western blotting. (E) HeLa cells were transfected with control siRNA or FAF1 siRNA and replated on coverslips for 72 h. Cells were treated with vehicle or Y-27632 (10 μM) for 6 h and then stained with F-actin (phalloidin) and β-catenin. Scale bar, 10 μm.