Figure 3

Apoptotic signaling was regulated by ANT1 transfection. A. ANT1 transfection induced Akt inactivation, reduced Bcl-xL expression, and Bax induction. Cells were transfected with pcDNA-ANT1 for 12 hours, and then total or nuclear extracts were prepared and Western-blotted using anti-phospho-Akt, anti-Akt, anti-Bcl-xL, anti-Bax, or anti-α-tubulin antibodies. Immunoblots were visualized by enhanced chemiluminescence. B. Mitochondrial membrane potential disruption induced by ANT1 transfection. Cells were transfected with pcDNA-ANT1 for 12 hours and then stained with DiOC₆. Mitochondrial membrane potentials were quantified by flow cytometry. C. Caspases-9 and -3 activation induced by ANT1 transfection. Cells were transfected with pcDNA-ANT1 for 24 hours. Cytoplasmic extracts were then prepared and Western-blotted using anti-caspase-9, anti-caspase-3, or anti-α tubulin antibodies. Immunoblots were visualized by enhanced chemiluminescence.