Figure 5.

Effect of IFNγ and TNFα costimulation on cytochrome c, Bcl-2, and Bax levels in MC3T3-E1 cells. Cells were cultured as described in Figure 4 and treated with 10 ng/ml IFNγ and 5 ng/ml TNFα. Samples were collected at various time points and fractionated into cytosolic and mitochondrial (Mito) fractions, of which 20 μg protein was subjected to 12% SDS-PAGE and immunoblotted with antibodies to cytochrome c. The mitochondrial fraction was also analyzed by western blot for Bcl-2 and Bax. Results are representative of three independent experiments.