Accuri plus software

Treated or untreated cells were collected and fixed in 70% ethanol for 1 h at 4 °C. For cell cycle analysis analysis, fixed cells were collected by centrifugation, washed with ice cold 1X PBS followed by staining with PI cell cycle reagent (BD). Cell cycle analysis was carried out by counting 5000 events and acquired data was analyzed using BD Accuri Plus software (BD).
For flow cytometry analysis of CSC markers, cells were collected by centrifugation, washed with ice cold 1X PBS followed by staining for CD24-PE (BD), CD44-APC (BD), CD-117-PerCp(BD), CD146-FITC (BD). Cells were analyzed in BD Accuri Plus Flow Cytometer (BD). For 20,000 events, percentages of positive populations were determined by using BD Accuri Plus software (BD).

Treated or untreated cells were washed with PBS and stained with AnnexinV-FITC/PI kit(Life Technologies, UK) according to the manufacturer’s protocol. In short, cells were incubated with AnnexinV and PI for 15 min in the dark and then analyzed at a flow cytometer using BD Accuri Plus software (BD).