Cell Apoptosis and Cell Cycle Assays

In brief, transfected cells were harvested, and washed with PBS (Invitrogen), followed by fixation with ethanol for 1 h. After measurement with FACSan flow cytometry (BD Biosciences) with 5 μL Annexin (V-fluorescein isothiocyanate) V-FITC/Propidium Iodide (PI) (Beyotime, Nantong, China), apoptosis cells were analyzed with Quest software (BD Biosciences). For cell cycle assay, after treatment with ethanol and PBS (Invitrogen), cells were added to RNaseA and PI for 30 min at room temperature. Finally, FACSan flow cytometry (BD Biosciences) and ModFit LT 3.2 software (Verity Software House, Topsham, ME, USA) were utilized to analyze the percentages of cells in the G0-G1 and S phases.

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