Fig. 3

Cytosolic localization of cytochrome c following dual treatment with FCCP and TRAIL indicates induction of apoptosis in KGN cells. Cytochrome c release was assayed via immunocytochemistry to detect induction of apoptosis. Following treatment with FCCP and/or TRAIL, cells were fixed and immunolabeled with antibodies against (a) the mitochondrial marker ATPB (green) and (b) cytochrome C (red). In addition, cells were also labeled with DAPI to visualize the cell nuclei and establish random fields of view for imaging. c Representative image indicating a cell (white arrow) scored as having released cytochrome C due to the loss of colocalization with ATPB. d Vehicle treated and TRAIL treated cells contained a small percentage of cells with released cytochrome C whereas both FCCP treated and FCCP and TRAIL treated cells showed significant increases in the percentage of cells exhibiting released cytochrome C (59.05 ± 5.39%, P < 0.05, 58.17 ± 15.65%, P < 0.05, respectively. Differing letters indicate significance). Scale bar indicates 50 μm