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An Annexin V-FITC Apoptosis Detection Kit (Beyotime, Shanghai, China) was used to conduct the apoptosis assay. Cells treated with KPT-335 for 48 h were collected from the culture medium and washed three times with PBS. Next, 5 μL of FITC, 5 μL of propidium iodide (PI) (50 μg/mL), and 200 μL of Annexin V binding buffer (1 ×) were added to the mixture. Then, the cell suspension was incubated in the dark at room temperature for 15 min. Apoptosis analysis was performed using FACSCalibur (BD Biosciences, San Diego, CA, USA).
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