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To study cell apoptosis, propidium iodide (BioVision, USA) double staining and Annexin V-fluorescein isothiocyanate staining were performed according to the manufacturer’s instructions. The cell apoptosis was detected by flow cytometric analysis (BD FACSCanto II Flow, Beckman-Coulter, USA). The cells in the upper left area are cells after necrosis, which was reduced to 0% by optimizing operations, such as the speed and time of centrifugation, selection of apoptosis kit, or the door of FSC/SSC.
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