Annexin 5 fitc and pi apoptosis kit

Cell apoptosis was determined by flow cytometric analysis using Annexin V-FITC and PI apoptosis kit (BD Biosciences, Franklin Lakes, NJ, USA). After treatment, cells were re-suspended in binding buffer, stained with 10 μL of Annexin V-FITC and 10 μL of PI, and then incubated at room temperature for 15 min in the dark. The stained cells were analyzed by a flow cytometer (BD Biosciences). The apoptotic cells were expressed as a percentage of the total number of cells.

Apoptotic cells were identified using Annexin-V/PI staining analyses. Briefly, cells were respectively seeded into 6-well plates and treated with IHZ-1 in different concentrations (1, 5, and 10 µM) or IHZ-1combine with chloroquine (10 μM) for 24 h. Cells were then harvested and washed using PBS. The surface levels of phosphatidylserine were quantitatively detected using Annexin V-FITC and PI apoptosis kit according to the manufacturer’s instructions (BD Biosciences, San Jose, CA, USA). The stained cell number were analyzed by flow cytometry (Accuri C6, BD Biosciences, Franklin Lakes, NJ, USA) within 1 h.