Annexin 5 allophycocyanin apc 4 6 diamidino 2 phenylindole dapi double staining kit

With the detection of apoptosis by flow cytometry, an Annexin V-allophycocyanin (APC)/4′,6-diamidino-2-phenylindole (DAPI) double-staining kit (Thermo Fisher Scientific) was used to analyze cellular apoptosis. Cells were seeded in 6-well plates (5 × 10⁵ cells/well) and then digested with trypsin (Gibco trypsin-EDTA; Thermo Fisher Scientific), washed with phosphate buffered saline (PBS) three times, suspended in 500 μL of binding buffer, and then incubated with 5 μL of fluorescein isothiocyanate (FITC)-conjugated Annexin V and 3 μL of propidium iodide (PI) for 15 min at room temperature in the dark. The stained cells were detected using the BD FACSAria II flow cytometer (BD Biosciences, Hercules, CA, USA).