Figure 10

Detection of CORT-induced apoptosis of mLTC-1 cells. A. mLTC-1 cells were incubated with either 100 nM CORT or 100 ng/mL CsA plus 100 nM CORT. The Leydig cells treated with a vehicle (Dimethyl Sulfoxide, DMSO) served as control. After 12-h incubation, the mLTC-1 cells were analyzed by FACS with annexin-V/PI double staining. The cells in the right lower quadrant were designated apoptotic (propidium iodide (PI)-negative/annexin V-fluorescein isothiocyanate-positive), the cells in the left lower quadrant were designated alive (PI-negative/annexin V-FITC-negative), the cells in right upper quadrant were designated dead (PIpositive/annexin V-FITC-positive), and the cells in left upper quadrant were designated damaged (PI-positive/annexin V-FITC-negative). B. The mLTC-1 cells treated with CORT showed increased frequencies of apoptotic labeling compared with the control. The Leydig cells treated with CORT plus CsA showed decreased frequencies of apoptotic labeling compared with the cells treated with CORT alone. * denote a significant difference compared with control at P < 0.05. ** denote a significant difference compared with cells treated with CORT at P < 0.05.