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The cells (15105) were then resuspended in 500 l binding buffer, mixed with 5 l annexin V-FITC (KenGen Biotech Co., Ltd., Nanjing, China) and 5 l propidium iodide sequentially, incubated at room temperature in the dark for 515 min, and detected by flow cytometry (Beckman Coulter, Inc., Brea, CA, USA) within 1 h