Fig. 4

Ovarian stromal fibrosis in PCOS mice was reduced by NR supplementation. A Representative images of ovarian collagen content in three groups of mice measured by PSR staining. Scale bar, 500 μm. B Quantitative analysis of fibrotic area in ovarian sections from three groups of mice (n = 8 to 13 slices for each group). C Representative images of ovarian fibrosis in the three groups of mice measured by immunohistochemistry. Scale bar, 500 μm. D Quantification of immunohistochemical staining intensity of ovarian sections from the three groups of mice (n = 16 slices for each group). E Transcriptional levels of Sirt1, Sirt3, Tgfb1, and Smad3 genes in the ovaries were measured by qRT-PCR (n = 3 replicates from 3 mice). F Diagram of the isolation of ovarian stromal cells. G, H qRT-PCR was conducted to validate the purity of the isolation. I NAD.⁺ levels in ovarian stromal cells (n = 3 replicates of cells pooled from 4 mice). J Transcript levels of Col1a1, (K) Col1a2, and (L) Col3a1 genes in ovarian stromal cells detected by qRT-PCR (n = 3 replicates of cells pooled from 4 mice). Data are presented as the mean ± SD. *P < 0.05, **P < 0.01, and ***P < 0.001 by one-way ANOVA compared to DHEA-treated mice (B, F-I) or Student's t-test (D, E)