After transfection for 48 h, the cell cycle detection kit (KGA512, KeyGEN, China) and the apoptosis detection kit (KGA106, KeyGEN) were used to measure cell cycle distribution and apoptosis, respectively [22 (link)]. The cells used to detect cell cycle were washed with PBS and collected by centrifugation (310 g, 5 min). They were then fixed with 70% ethanol overnight at 4°C, rinsed, and suspended in 0.5 mL solution including propidium iodide (PI) and RNaseA (PI:RNaseA = 9:1). After incubation for 45 min at room temperature (RT), they were analyzed by FCM (C6, BD Sciences, USA).
The cells used to detect cell apoptosis were resuspended. Then, the Annexin V-FITC solution (5 μL) was added to cells and mixed, and then PI (5 μL) was added. After reaction at RT for 10 min in the dark, the apoptosis rate was assessed via FCM.
The cells used to detect cell apoptosis were resuspended. Then, the Annexin V-FITC solution (5 μL) was added to cells and mixed, and then PI (5 μL) was added. After reaction at RT for 10 min in the dark, the apoptosis rate was assessed via FCM.
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