Flow cytometry to measure apoptosis and cell cycle progression following cisplatin treatment

Cells were cultured with increasing concentrations of cisplatin and incubated for 24, 48, and 72 h. PE-annexin V and 7-AAD (1 μl each) were mixed and added to cells. This was incubated for 15 min at room temperature in the dark. Next, 400 μl of ice-cold binding buffer was added and mixed gently before the cell preparations were examined by flow cytometry. Flow cytometry was then used to analyze the rate of apoptosis. For cell cycle analysis, cells were treated with cisplatin for 48 h. 1 μl RNA enzyme and 10 μl Triton X-100 were added to cells at room temperature for 30 min. Next, 1 μl PI was added and incubated at 4 °C for 30 min in the dark. Finally, flow cytometry was performed.