Cell Cycle and Apoptosis Analysis

The cell cycle distribution and apoptotic cell population were scrutinized as described previously (23 (link)). In brief, for the cell cycle, collected cells were fixed with 70% ethanol (Sangon Biotech, Shanghai, China) and stained with 0.5 ml propidium iodide (PI) (Beyotime Biotechnology, Shanghai, China) for 30 min. For cell apoptosis analysis, collected cells were stained with 5 μL Annexin V-PE and 5 μL 7-AAD for 10 min (Vazyme, Nanjing, China). The cells were examined using flow cytometry (BD, New Jersey, USA). The cell cycle data were analyzed with FCS Express Launcher software (De Novo Software), and the cell apoptosis results were analyzed with FlowJo software.