Apoptosis and Cell Viability Assay

HEK293 cells were cultured in DMEM supplemented with 10% fetal bovine serum, penicillin (10 U/mL), and streptomycin (100 μg/mL), and treated with 0.3 or 1 μmol/L of {"type":"entrez-protein","attrs":{"text":"SCH79797","term_id":"1052762130","term_text":"SCH79797"}}SCH79797 or 0.1% of DMSO (v/v) as vehicle for 3, 6, or 24 hours. After this period, cells were lysed with RIPA lysis buffer, and the ratio of cleaved Casp3 (caspase‐3; 1:1000; Cell Signaling) to Casp3 (1:1000; Cell Signaling) was evaluated by western blot analysis. As a positive control, we administrated staurosporine 1 μmol/L (Abcam) to cells to induce apoptosis and collected cells 4 hours later. In a cell‐viability assay, HEK293 cells were cultured in DMEM supplemented with 10% fetal bovine serum, penicillin (10 U/mL), and streptomycin (100 μg/mL), and treated with 0.1, 0.3, or 1 μmol/L of {"type":"entrez-protein","attrs":{"text":"SCH79797","term_id":"1052762130","term_text":"SCH79797"}}SCH79797 or 0.1% of DMSO (v/v) as vehicle for 3 or 24 hours. After this period, cells were detached from the culture dish using Trypsin‐EDTA (Wako), and then trypan blue solution (Wako) was added. The number of whole cells and stained cells were counted with a TC20 automated cell counter (Bio‐Rad), and the survival rate was calculated.