Fig. 4.

Modulation of CTGF expression by Bcl-2 through YAP in M14 cells. A Western blot analysis of YAP phosphorylation, pYAP (S127), and YAP protein in total protein extracts of control (Control) and Bcl-2 overexpressing (Bcl-2/6) clones. B) Western blot with relative densitometric analysis expressed as “nuclear/cytosol ratio” of YAP protein expression in the nuclear (nucleus) and cytoplasmic (cytosol) extracts of Control and Bcl-2/6 clones. α-tubulin and histone H3 were used as cytoplasmic and nuclear loading control, respectively. One representative western blot analysis out of three with similar results is reported. C Western blot with relative densitometric analysis and D qRT-PCR analyses of YAP and CTGF expression in Bcl-2/6 cells transiently transfected with siRNA SMARTpools targeting YAP (si-YAP) or with control siRNA (si-Ctrl). Data represent ratio (mean ± standard deviation) of mRNA expression normalized to β-actin in silenced cell condition versus controls from three independent biological repeats. E Western blot and densitometric analyses of YAP and CTGF proteins expression in Bcl-2/6 cells in the presence or absence of Verteporfin, VP (1 µM, 24 h). A-C, E Representative images of three independent experiments with similar results. α-tubulin and vinculin are shown as loading and transferring control. F ELISA analysis of CTGF protein levels in Conditioned Medium (CM) derived from Bcl-2/6 cells treated with VP. CTGF levels were normalized to the number of adherent cells and expressed as ratio (mean ± standard deviation) of treated cells over control in three independent experiments. B-F Statistical analysis was performed applying unpaired two-tailed student’s t test, *p < 0.05