Figure 5

Western blotting of cytochrome c released from mitochondria. Jurkat cells (5 × 10⁵/ml) were treated with Pep27anal2 at 12.5 μM for 1 h, 2 h, and 4 h, and at 62.5 μM for 4 h. Mitochondria were pelleted by centrifugation, and the supernatants retained. As a control, mitochondria were treated with 0.1% (vol/vol) SDS, which induces cytochrome c release from mitochondria. Protein from each fraction was subjected to SDS/PAGE for Western blot analysis using an anti-cytochrome c antibody. 1, the result of analyzing 50 μg of cytosol after adding no Pep27anal2; 2, 50 μg cytosol after 12.5 μM for 1 hr; 3, 50 μg cytosol after 12.5 μM Pep27anal2 for 2 hr; 4, 50 μg cytosol after 12.5 μM Pep27anal2 for 4 hr; 5, 50 μg cytosol after 62.5 μM Pep27anal2 for 4 hr; 6, 50 μg mitosol after 0.1% SDS; 7, 25 μg mitosol after 0.1% SDS.