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Figure 2 | Cell Communication and Signaling

Figure 2

From: Chk1 activity is required for BAK multimerization in association with PUMA during mitochondrial apoptosis

Figure 2

Effect of Chk1 inhibitor on cytochrome c release from mitochondria and caspase activation. (A) Mitochondria were isolated from HCT116-WT BAK cells and HCT116-WT BAK cells treated with Chk1 inhibitor (SB-218078) for 30 min. Mitochondria were then incubated with increasing concentrations (0.5-1 ng/μl) of recombinant tBid (R&D) at 37°C for 30 min. Mitochondria were then separated into pellet and supernantant fractions by centrifugation and western blotted for Cyt c ab (BD pharmingen). An equal amount of mitochondria not treated with tBid was used as input. (B) Caspase activation was monitored by immunoblotting with anti cleaved caspase3 antibody using total cell extracts from HCT116-WT BAK cells treated with or without Chk1 inhibitor for 30 min following UV. (C) Similarly caspase3 activation was analysed in HT1080 and HT1080-Chk1D-A (Chk1 D130A transfected in HT1080 cells) by western blotting with anti cleaved caspase 3 antibody. Actin represents equal loading.

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Cell Communication and Signaling

ISSN: 1478-811X