The cells were detached with 0.25% trypsin (90058, Thermo Scientific). After detachment, 100 μL binding buffer was added to prepare cell suspension (1 × 106 cells/mL). Annexin V-fluorescein isothiocyanate (FITC, 40302ES20, Yeasen Biotechnology Co., Ltd., Shanghai, China) and propidium iodide (PI) were added sequentially for a 5-min incubation in darkness at ambient temperature. The FC500MCL flow cytometer (FACS Canto II, BD Biosciences, San Jose, CA, USA) system was applied to assess apoptosis.

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