Figure 3.

Bax is integrated into the mitochondrial membrane in response to grB and Ad. Jurkat targets were treated either with nothing (control [ctrl]), grB alone, Ad alone, or grB and Ad (grB/Ad) in the presence or absence of 100 μM zVAD-fmk for 3 h at 37°C. After incubation, the plasma membranes of cells were rendered permeable with digitonin. The permeable cells were then centrifuged to separate the pellets (containing mitochondria) from the supernatants (cytosol). Pellets were subjected to an alkaline extraction with 0.1 M Na₂CO₃ for 30 min on ice followed by centrifugation. The pellets and the prealkaline extraction supernatants were resolved on 15% polyacrylamide gels. Proteins were transferred to nitrocellulose and the blots were probed with polyclonal anti-Bax antibodies. These data are representative of three independent experiments.