5-azacytidine and decitabine induce expression of active caspase-8 and active caspase-3 in neoplastic MCs. (A) HMC-1.1 and HMC-1.2 cells were incubated in control medium or in medium containing 5-azacytidine (5μM) or decitabine (5μM) at 37°C for 96 hours. Then, Western blotting was performed using antibodies against cleaved caspase-8 and cleaved caspase-3. To confirm equal loading, a polyclonal antibody against β-actin was applied. (B) Cord blood progenitor-derived normal MCs and HMC-1 cells were cultured in the presence of control medium (left panels), 5-azacytidine (5μM; middle panels), or decitabine (5μM; right panels) at 37°C for 96 hours. Then, cells were analyzed by annexin V/PI staining. Results show the percentage of annexin V/PI-positive cells determined by flow cytometry. (C) HMC-1.1 cells and HMC-1.2 cells were incubated in control medium (Co) or in medium containing 5-azacytidine (5μM) or decitabine (5μM) at 37°C for 96 hours. After incubation, the presence of apoptotic cells was determined by TUNEL assay.