Fig. 3

Quercetin reduces the Cy-induced phosphorylation of PI3K/Akt/Foxo3a pathway proteins in the ovary. a Western blot analyses were used to evaluate ovaries from 8-week-old mice removed at 24 h after a single dose of Cy (75 mg/kg) or an equivalent volume of PBS with or without quercetin (Que1: 20 mg/kg; Que2: 40 mg/kg). Comparisons of the concentrations of phosphorylated and total Foxo3a, Akt, mTOR, and rpS6, using β-actin as a loading control. Fold-change values for ratios for each protein are listed below each set of bands. b Ratios of phosphorylated/total protein are summarised in a bar graph. Experiments were repeated three to five times with similar results; three to five ovaries were pooled per result, with a total of 45 animals. The data are presented as the mean values ± SEM. (***P < 0.001 compared with the Cy group; # P < 0.05, ## P < 0.01, ### P < 0.001 compared with the control group). One-way ANOVA followed by Tukey’s tests was used for statistical analyses. c-d Immunohistochemical staining for Foxo3a and Ki67 demonstrates that quercetin inhibited Cy-induced primordial follicle overactivity and short-term growth. A representative image from each group is shown. c immunohistochemical staining for Foxo3a shows the localisation of Foxo3a in each group. Arrows indicate primordial follicles. d Ki67 staining of ovary tissues was used to evaluate cell proliferation. Black frames indicate primary follicles. Scale bars, 20 μm