Figure 6.

Bax insertion into mitochondrial outer membranes is not affected by ionic strength. (A) Mitochondrial suspensions (960 μg mitochondrial protein/mL) were treated with 50 nM Bax and 240 nM tBid for increasing amounts of time (10, 20, and 30 min) in either 10 mM or 90 mM KCl buffer. Then the mitochondria were centrifuged and supernatant assayed for AK release. The pellets were subjected to carbonate treatment and probed for Bax insertion by Western blot. The carbonate treatment Western blots were performed as described in the Materials and Methods in the Supporting Material. VDAC served as loading control. (B) Extent of AK release at 10 (solid line) and 90 mM KCl buffer (dashed line) for time points corresponding to those in A.