Fig. 5.

Effects of H2O2 and/or melatonin on apoptosis of granulosa cells. (A) Granulosa cells were incubated with H2O2 (0.1, 1, 10 mM) for 2 h. (B) Granulosa cells were incubated with H2O2 (10 mM) in the presence or absence of melatonin (100 μg/ml) for 2 h. Apoptosis of granulosa cells was assessed by nuclear fragmentation using DAPI staining. Apoptotic cells were identified by condensation and fragmentation of the nuclei. Apoptosis was quantified by calculating the percentage of apoptotic nuclei in a total of 200 nuclei in each group. Data are shown as the mean ± SEM of three independent incubations. a, P < 0.05 vs. control, and b, P < 0.05 vs. the other groups (Kruskal-Wallis H-test and Mann-Whitney U-test with Bonferroni correction).