FIG. 6.

HGTD-P induces mitochondrial PT with resultant cytochrome c release and caspase 9 activation. PC-3 cells were transiently transfected with 1 μg of mock vector or pcDNA-HGTD-P in the presence or absence of 20 μM cyclosporine (CsA) or 100 μM bonkrekic acid (BA) or cotransfected with 1 μg of pcDNA-HGTD-P and 1 μg of pcDNA-Bcl-2. (A) After 24 h of transfection, cells were incubated with 400 nM potential-sensitive Mitotracker Red CMXRos, and then MTP was determined by flow cytometry. (B) After 36 h of transfection, cells were harvested, fractionated, and subjected to Western blotting for cytochrome c. Cyto., cytosolic compartment; Mito., mitochondrial compartment. (C) Cells were harvested at the indicated time points, and the caspase 9 assay was performed. The caspase activities of mock-transfected cells were arbitrarily set at 1. The increase in caspase activity is presented.