CAOV3 and SKOV3 cells were digested with trypsin without EDTA. The cells were harvested and diluted to a density of approximately 1×106 cells/ml after 24 h transfection, then washed twice with ice-cold phosphate-buffered saline. Subsequently, the cells were incubated in the dark, at room temperature for 15 min with 5 µL phycoerythrin (PE) Annexin V, 5 µL 7-aminoactinomycin D (7-AAD) and 500 µL 1×binding buffer from the Annexin V-PE/7-AAD apoptosis kit (KeyGEN Bio TECH Corp, China) according to the manufacturer's protocol, then they were analyzed by flow cytometry. Cells undergoing early apoptosis only bound to PE Annexin V, whereas cells that bound to PE Annexin V and 7-AAD were in the late stages of, or had already undergone, apoptosis.

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