Fig. 3

CDDP induces p-p53 (ser15)-Phb1-Bak interaction in chemosensitive but not in chemoresistant gynecologic cancer cells. A A2780s (chemosensitive OVCA cells) and A2780cp (chemoresistant OVCA cells) were treated with CDDP (0–10 μM, 6 h). Protein contents of Phb1, p-p53 (ser15), Bak, and GAPDH were examined by WB. Protein-protein interaction was determined by immunoprecipitation (IP)-WB. Cell lysates were immunoprecipitated with anti-IgG (control; lanes 1) or anti-Bak antibody. Bak immunoprecipitates were immunoblotted [IP: anti-Bak, WB: anti-Bak, −Phb1, −p-p53 (ser15)]. B CDDP-induced Phb1-dependent p-p53 (ser15)-Phb1-Bak interaction. OV2008 (chemosensitive CECA cells) were forced expressed with Phb1 siRNA (0–100 nM, 24 h; scrambled siRNA as control) and then treated with CDDP (0–10 μM, 6 h). Contents of Phb1, p-p53 (ser15), Bak and GAPDH were examined by WB. Cell lysates were immunoprecipitated with IgG (control; lanes 1) or Bak antibody. Protein-protein interaction was determined by IP-Western. Bak immunoprecipitates were immunoblotted [IP: anti-Bak, WB: anti-Bak, −Phb1, −p-p53 (ser15)]. Results are expressed as mean ± SEM (n = 3) and analyzed by 2-way ANOVA and Bonferroni post-hoc test. [*p < 0.05, **p < 0.01, ***p < 0.001 (versus DMSO (A) and CDDP in absence of Phb1siRNA (B)); n = 3]