Fig. 4.

Bcl-2 siRNA abolishes the protective activity of ANG. (A) Western blotting analysis of the protein level Bcl-2 in vector control and in Bcl-2 specific shRNA transfectants. The bar graph at right is the relative density of Bcl-2 with β-actin. (B) AIF protein level in the nuclear fractions extracted from vector control and Bcl-2 shRNA transfectants cultured in serum-free medium with or without ANG for 24 h. The left panel is Western blotting film with an anti-AIF IgG. The right panel is relative intensity of AIF with Histone H3 as the normalization control. (C) The protein level of cleaved PARP-1 in the total cell lysates from vector control and Bcl-2 shRNA transfectants incubated with or without ANG for 16 h. The left panel is Western blotting film with anti-cleaved PARP-1 IgG. The right panel is the relative intensity of cleaved PARP-1 with actin as the normalization control. (D) The protein level of pro-caspase-3 and activated caspase-3 in total cell lysate from vector control and Bcl-2 shRNA transfectants incubated with or without ANG for 2 h. The left panel is Western blotting film with anti-caspase-3 IgG. The right panel is the relative density of active caspase-3 vs pro-caspase-3. The Western blotting images were from a representative experiment. The bar graphs shown are mean ± SD of three independent experiments.