Figure 5

From: Desmoglein 3 promotes cancer cell migration and invasion by regulating activator protein 1 and protein kinase C-dependent-Ezrin activation

Figure 5

Dsg3 activates the ERM proteins including Ezrin at the Thr567 residue. (a) Western blotting analysis with the indicated antibodies showed an enhanced phosphorylation of pERM in A431 cells with high Dsg3 expression levels. The representative blots are shown on the left and bar charts on the right (experiment n>6, mean±s.e.m., *P<0.05). (b) C7 cells transfected with either the scrambled control or Ezrin- or moesin-specific small interfering RNA for 2 days before protein extraction followed by western blotting. Total protein samples (10 μg per lane) were resolved in 3–8% SDS–polyacrylamide gel electrophoresis that showed a good separation of Ezrin/radixin (81 kD) and moesin (77 kD). There was a good knocking down for Ezrin but less effect for moesin, and the bar chart summarizes the data of four independent blots (mean±s.e.m., ***P<0.001). (c) PLA assay using antibody pair for pERM/Ezrin showed significantly increased PLA signal in D3 cells compared with Vect cells (mean±s.d., *P<0.05). (d) Confocal images of A431 cells showed rounded cells coupled with pronounced membrane protrusions (enlarged on the left) in cells with high Dsg3 levels when compared with control cells or cells with low Dsg3 levels in the same sample. Scale bar, 20 μm.