2.5　Cell apoptosis assay

Cells were cultured in 12-well culture plates with different treatments for 4h，then washed twice with cold PBSand resuspended in binding buffer to a concentration of 1×106 cells/mL.Next，100μL of the resuspended cells was transferred to a 5-mL tube，5μL of Annexin V-FITC and 5μL PI were added，and the cells were gently vortexed.After 15min of incubation at room temperature，400μL of binding buffer was added to each tube and cells were analyzed by flow cytometry within 30min.(https://www.daowen.com)