Mitochondrial targeting and membrane insertion of Bax are not affected by N-terminal cleavage or Asp33Ala mutation. Inducible Bax expression was performed in the presence of 40 μM ALLM for the indicated times. The heavy membrane was isolated, and alkali extraction was performed to determine the percentage of the alkali-resistant (integral, nonextractable) Bax contained in the mitochondrial membrane as described in “Materials and methods.” Intermembrane cytochrome c was used as extractable positive control, and inner membrane prohibitin was used as nonextractable negative control.