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Fig. 1. Limited role of caspase 3 and 7 activation in EGF-induced cell death. (A) A431/ErbB2 or SKBr3/EGFR cells were grown until 70-80% confluency and then treated with EGF (20 ng/ml) or vehicle in the presence or absence of caspase inhibitors (Z-VAD-FMK, 30 µM; caspase inhibitors 3 and 9, 20 µM) for the indicated times. Staurosporin-treated cells were used as a positive control. The cells were lysed in CHAPS buffer as indicated in Materials and Methods. The levels of activated caspases were detected by western blotting using antibodies against cleaved caspase 3 and caspase 7. (B) MCF7 cells, which are devoid of caspase 3, were transfected with EGFR*GFP and ErbB-2*RFP constructs and stable cell lines were selected. The cells were incubated with or without EGF for 4 days and images were prepared using fluorescence microscopy.
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