2.5　Cell apoptosis assay

Apoptotic cells were analyzed by flow cytometry using PI-annexin V/FITC Apoptosis Detection Kit.MIN-6 cells were cultured in 12 wells，then washed twice with cold PBSand resuspended in 100μL binding buffer at a concentration of 106 cells/mL.This is followed by adding 1μL of annexin V-FITC，1μL PI and 400μL binding buffer to each tube，gently vortexing the cells and incubating for 10min at room temperature，and finally analyzing by flow cytometry within 30min.(https://www.daowen.com)