Figure 5.

Prolonged maintenance of gctBid is associated with cyt c release after ABT-737 treatment up to 18 h after initiation of the GraB death signal. HeLa and HeLa-Bcl-2 cells were treated with Pfp/GraB in the presence of Q-VD-OPh (QVD; 20 μM). C20 (10 μM) was either added to HeLa cells (lane marked as C20*) together with the Pfp/GraB (at 0 h) or to the Bcl-2 cells after 2 h (lanes marked as C20^#). ABT-737 (500 nM) was added to Bcl-2 cells as indicated after a further 15 h (t=17 h) and the cells were fractionated at 17.5 h. Mitochondrial fractions were examined by Western blot analysis for the presence of gctBid (upper panel) and cyt c loss from the mitochondria (middle panel). cyt coxidase (CCO) levels indicated equal protein loading (lower panel). Results are representative of three similar experiments