Figure 8

SiRNA-mediated knockdown of SIRT3 abolishes the protective effects of melatonin in granulosa cells. SIRT3 was knocked down by SIRT3 siRNA transfection as described in the Materials and Methods section.

(a) Mitochondrial ROS, (b) SIRT3 activity, (c) Cell viability, (d) GSSG/GSH ratio, (e) 8-OHdG, (f) Apoptosis ratio. (g) The expressions of SIRT3, SOD2, CAT and the nuclear translocation of FoxO3a were examined using immunoblotting. (h) Granulosa cells were harvested for ChIP analysis and the FoxO3a binding affinity was measured by real-time PCR assay. The values are presented as the mean ± SD of the results from three independent experiments. *p < 0.05, **p < 0.01 versus the Control siRNA + H₂O₂ group; ^#p < 0.05, ^##p < 0.01 versus the Control siRNA + H₂O₂ + melatonin group; ^$p < 0.05, ^$$p < 0.01 versus the SIRT3 siRNA + H₂O₂ group. For ChIP assay, *p < 0.05 versus the si-Scramble + H₂O₂ group; ^##p < 0.01 versus the si-Scramble + Mel + H₂O₂ group.