Figure 5.

The TM1 domain regulates inhibition by Bcl-x_L. A, Schematic representation of Bax and Bax 30–146. B, HEK cells transfected with Bax 30–146-GFP (1 μg) with or without Bcl-x_L (1 μg) and assessed for apoptotic nuclear damage after 15 hours of culture. Data are normalized for apoptotic damage triggered by GFP alone in both conditions. C, HEK cells were transfected with 1 μg GFP or Bax-GFP or Bax 30–146-GFP or Bax 30–105-GFP with or without 2 μg dominant negative caspase 9 (DNC9). Apoptotic nuclear damage was assessed after 15 hours of culture. Data is normalized to damage in cells transfected with GFP alone or GFP + DNC9 transfected cells. D, HEK cells transfected with Bax-GFP or Bax 30–146-GFP or Bax 30–105-GFP were harvested after 15 hours of culture and subjected to sub-cellular fractionation. The cytosolic (Cyto) and membrane/mitochondrial (Mito) fractions were assessed for GFP and Cox4 distribution by western blot analysis. WCL: whole cell lysate. Values in the anti-GFP blots are derived from densitometry analysis and indicate the amount of GFP detected in the specific fraction relative to the total signal in the mitochondria and cytosol fractions. The data are presented as mean ± SD derived from a minimum of three independent experiments. **p < 0.01; *p < 0.001.