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Cells were collected and centrifuged at 1500 rpm for 5 min. The cells were washed with ice-cold PBS and fixed with 70% ethanol overnight at −20 °C. The fixed cells were washed with PBS for 10 min and treated with RNAase A for 30 min followed by incubation with propidium iodide for 30 min at room temperature. The cell cycle in each specific sample were evaluated by the flow cytometer (FACS Calibur, BD Biosciences, USA) following the manufacture’s instruction.
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