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Annexin V-FITC/propidium iodide double staining was used to assess apoptosis and necrosis. According to the manufacturer's instructions of the Annexin V-FITC Apoptosis Detection Kit (R&D Systems, New Brighton, USA), samples were prepared for flow cytometric analysis. Viable cells were identified as Annexin-V−/PI−, early apoptotic cells as Annexin-V + /PI−, late apoptotic cells as Annexin-V + /PI +, and necrotic cells as Annexin-V−/PI +.
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