Fig. 5.

Regulation of Hippo pathway core proteins by Bcl-2 in cancer cells. A, B Western blot and relative densitometric analysis of LATS1 and MST2 proteins in A A375 cells transiently transfected with siRNA targeting Bcl-2 (si-Bcl-2) or control siRNA (si-Ctrl) and of LATS1, MST2 and MOB1 proteins in B A375 control (Control) and Bcl-2 overexpressing (Bcl-2/1) stable clones. The expression level of Bcl-2 protein in the stable clones was also verified. C Western blotting and densitometric analyses of phosphorylated LATS1 (pLATS1) and total LATS1 in A375 control and Bcl-2 overexpressing (Bcl-2/1) cells. D Western blot analysis of MST2 and relative densitometry from A375 control and Bcl-2 overexpressing (Bcl-2/1) cells in the presence or absence of MG132. E, F Western blot and relative densitometric analysis of LATS1 and MST2 proteins in E H460 cell s transiently transfected with siRNA targeting Bcl-2 (si-Bcl-2) or with control siRNA (si-Ctrl) and F in MDA-MB-231 cells transiently transfected with empty (pEmpty) or Bcl-2 (pBcl-2) expressing vectors. Bcl-2 protein expression was evaluated to validate overexpression efficiency. A-F One representative western blot analysis out of three with similar results is reported. HSP72/73 and α-tubulin were shown as loading and transferring control. Data represent ratio (mean ± standard deviation) of protein density normalized to HSP72/73 or α-tubulin in silenced or overexpressing cell conditions versus controls from three independent biological repeats. Statistical analysis was performed applying unpaired two-tailed student’s t test, *p < 0.05, **p < 0.01