Annexin V-FITC/PI Apoptosis Assay

An Annexin V FITC/PI Apoptosis Detection Kit (556547, Becton Dickinson, Franklin Lakes, United States) was used for the detection of cell apoptosis according to the manufacturer’s instructions. Briefly, cells in different groups were harvested by digestion with trypsin (25200072, Gibco, Grand Island, NY, United States), then washed twice with cold PBS, and resuspended in Binding Buffer. The cells were further stained with the mixture of 5 µL FITC Annexin V and 5 µL PI for 15 min at room temperature in the dark (Park et al., 2021 (link)). The apoptosis ratio was quantified with a Flow Cytometer (Beckman Coulter, Breya, California, United States).