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Seno et al. Cancer Drug Resist 2019;2:335-50 I http://dx.doi.org/10.20517/cdr.2019.01 Page 343

A B

C

D

E

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