Fig 2: MMINA upregulated protein expression levels of the key testosterone synthesis factors and related hormones in rat testes. (A) Effect on reproductive hormones; (i): testosterone, (ii): LH. (B) Effect of MMINA on transcriptional regulation of the steroidogenic hormone biosynthesis gene. rtPCR analysis of StAR (i), CYP11A1 (ii), and 3β-HSD (iii) mRNA expression. (C)-StAR, CYP11A1, and 3β-HSD protein content were determined by Western blot analysis. Values were expressed as mean ± standard deviation (n = 7). **, **** p < 0.05 and p < 0.0001 versus control, respectively, and ++, ++++ p < 0.05 and p < 0.0001 versus CDDP. Data were analyzed by one-way ANOVA followed by Tukey’s multiple comparison tests using graph pad prism version 9. STAR: steroidogenic acute regulatory protein, CYP11A1: P450 side-chain cleavage enzyme, HSD17B: 17β-hydroxysteroid dehydrogenase. Uncropped blots are presented in Supplementary File S3.

Fig 3: Statistical evaluation of serum testosterone (a), LH (b), and ucOC (c) levels of subjects sacrificed on the 4th day after EDS administration. ∗∗∗p < 0.0001; ∗∗p < 0.001; ∗p < 0.01.

Fig 4: Effects of Vitex nigundo on (A) LH (B) FSH (C) LH:FSH ratio in rats with letrozole induced PCOS. Data are presented as the Mean ± S.E.M. (n = 6), evaluated by ANOVA followed by the post hoc Tukey’s test. *P < 0.05, **P < 0.01, ***P < 0.001 vs. control; #P < 0.05, ##P < 0.01, ###P < 0.001 vs. LTZ group. The inter and intra assay variation (CV) was < 10% for all parameters