2.8. Mitochondrial membrane potential (DWm) detection
Change in DWm was quantitatively determined by staining the cells with the
cationic dye, JC-1, following the manufacturer’s instructions (Sigma, St. Louis,
USA). Briefly, after 48 h treatment, cells were washed with PBS and incubated with
medium containing 2.5 lg/ml of JC-1 staining reagent at 37 C for 20 min followed
by a wash with ice-cold 1 JC-1 staining buffer. The stained cells were analyzed by
FACScan flow cytometry equipped with CellQuest software. A total of 20,000 cells
were analysed for each treatment.