Annexin-V/PI apoptosis assay

HEK293T cells were transfected with Bqu-BepC tagged with Strep and control plasmid (no gene insertion) for 24 h, or treated with 25 μM CCCP (Carbonyl cyanide m-chlorophenyl hydrazine, apoptosis inducer) for 12 h as positive control. Cell samples were trypsinized, collected and centrifuged at 2000 rpm to remove the supernatant. Next, cells were resuspended by adding binding buffer gently. The apoptosis and necrosis were subsequently detected using an FITC Annexin-V apoptosis detection kit according to the manufacturer’s instructions (Yeasen, China). Apoptotic cells stained with FITC Annexin-V and propidium iodide (PI) were analyzed using a BD FACSCalibur flow cytometer.