Flow cytometry assays for apoptosis analysis by Annexin V and PI double-staining

Evaluation of apoptotic cells was conducted by Annexin V and PI double-staining, and detected by flow cytometry. In brief, QGY-7703 and Bel-7402 cells were plated into 6-well plates at a density of 2×10⁵ per well and treated with DMSO or various concentrations of asiaticoside (1, 3, and 10 μM), followed by incubation for 48 h. Subsequently, cells were harvested and incubated with Annexin V (5 μL of 100 μL binding buffer) and PI (5 μL of 100 μL binding buffer) for 30 min. Then, the cells were washed and resuspended in 200 μL of binding buffer. A BD FACS Verse flow cytometer was used to detect apoptotic cells. FlowJo software was used to analyze the results.