fig3

Figure 3. Involvement of caspases in the apoptosis of miPS-LLCcm cells. A: Expression levels of procaspases during daunorubicin-induced cell death of miPS-LLCcm cells. miPS-LLCcm were treated with 100 nmol/L daunorubicin for the indicated periods. Whole cell lysates were prepared and analyzed by western blotting; B: detection of processed caspases in cells attached or detached to the dish. After daunorubicin treatment, floating cells were concentrated by centrifugation, and whole-cell lysates were prepared. Cleaved caspase-3, procaspase-9, and procaspase-7 levels were detected; C: confirmation of whole proteins in detached cells. Cell lysates were subjected to SDS-PAGE and stained with CBB; D: suppression of cleavage of PARP-1 by Z-VAD-FMK. Cells were treated with 100 nmol/L daunorubicin in the presence of the pan-caspase inhibitor, Z-VAD-FMK. Cleaved PARP-1 was analyzed by western blotting; E: suppression of DNA fragmentation by Z-VAD-FMK. DNA fragmentation in cells treated with daunorubicin in the presence of 50 μmol/L of Z-VAD-FMK were analyzed