Fig. 2

The regulating effects of p120 in E-cadherin endocytosis and epithelial gap formation. MLE-12 cells were transfected with p120 siRNA or p120 cDNA. After 48 h posttransfection, cells were exposed to 20 % cyclic stretches for 2 h. (a) Successful transfection was detected by western blot. (b, d) The effect of p120 siRNA and p120 cDNA transfection on E-cadherin degradation. Loss of p120 could significantly decrease the expression of E-cadherin. However, p120 overexpression reversed E-cadherin degradation. (c, f) Cytoplasm E-cadherin increased after p120 depletion while it was restored by p120 overexpression under cyclic stretch. (e) The association of p120 and E-cadherin was examined by immunoprecipitation after 2 h cyclic stretch in p120 cDNA transfection group. (g) Immunofluorescence and quantification data revealed the association of p120 and E-cadherin after stretch in p120 siRNA group and p120 cDNA group. Fixed Cells were incubated with anti-p120 and anti-E-cadherin primary antibody followed by Alexa-conjugated secondary antibody. (h) Immunofluorescence showed the gap formation in p120 siRNA group and p120 cDNA group. The F-actin was stained with Alexa 546 phalloidin (red). The nucleus were stained with DAPI (blue). Scale bars = 10 μm. *p < 0.05 versus control group, +p < 0.05 versus CS group. Experiments were repeated at least three times