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Cells were plated at 5 × 105 cells/dish into 60 mm dishes. After appropriate treatment and incubation, cells were collected with 0.25% trypsin, washed with cold PBS for twice, and stained using Annexin V-FITC/propidium iodide or PE/7-AAD for 15 min in dark at room temperature according to the manufacturers’ protocol. The rate of apoptotic cells was recorded using flow cytometry (BD Pharmingen USA). The experiment was repeated three times.
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