Regulation of Bax and bcl-2 expression by WT1 in immature rat GCs. Real-time RT-PCR analysis of (A) Bax and (B) bcl-2 mRNA levels in GCs transfected with increasing amounts of WT1 (−KTS) (10, 30, 100 ng/well) or WT1(+KTS) (100 ng/well). GAPDH was used to normalize each reaction. Values are calculated as fold changes relative to control (CT), and are expressed as the mean ± SD of three independent experiments, each performed in triplicate. CT, cells transfected with empty vector. *p < 0.05 compared to control. Western blot analysis of (C) Bax and (D) bcl-2 protein levels. Lysates were immunoblotted with anti-WT1 antibody (C-19; Santa Cruz Biotechnology, Inc.), anti-Bax antibody (B-9, SC 7480), or anti-bcl-2 antibody (C-2, SC 7382). Arrows indicate bands corresponding to WT1 (52 kDa), Bax (22 kDa), bcl-2 (26 kDa) and β-actin (42 kDa), respectively.