Skip to main content
. 2014 Oct 1;111(41):14782–14787. doi: 10.1073/pnas.1417253111

Fig. 1.

Fig. 1.

Inducible expression of Bim triggers mitochondria-mediated apoptosis. (A) A U2OS cell line with a stably transfected Bim transgene (U2OS_Bim) was treated with Dox (0.1 µg/mL) in the presence or absence of z-VAD (20 µM) for the times indicated. The same concentrations were used in all experiments. Cell viability was determined by measuring ATP levels using the Cell-Titer Glo kit. Following Dox treatment for the indicated times, the P15 fractions of U2OS_Bim cells were analyzed by Western blotting. (B) U2OS_Bim cells were treated with or without Dox for the times indicated. The S15 fractions of the cells were analyzed by Western blotting. (C) U2OS_Bim cells in the presence of z-VAD were treated with or without Dox for 4 h. Immunostaining was performed using anti-cytochrome c (green) and anti-Smac (red) antibodies or anti-translocase of the outer membrane 20 (TOM20) antibody. (Scale bars: 10 µm.) (D) U2OS_Bim cells were treated with Dox for the times indicated. The P15 fractions were cross-linked with 10 mM 1,6-bis(maleimido)hexane (BMH) where indicated and were analyzed by Western blotting.