Fig. 6: The role of FOXP1 in ovarian aging.

a, Network of regulatory TFs in three GC subtypes. Node size correlates with the number of edges positively. b, Network of regulatory TFs in five T&S subtypes. c, The correlation analysis of FOXP1 expression with age in hGCs and pT&S. n = 28–30 (Pearson correlation analysis, two-sided). d, FOXP1 expression in ovaries. Data are presented as the mean ± s.e.m. n = 10 for each group (one-way ANOVA). e, Western blotting of FOXP1 protein levels in ovaries. Data are presented as the mean ± s.e.m. n = 6 for each group (one-way ANOVA). f, SA-β-gal staining. Data are presented as the mean ± s.e.m. n = 5 for each group (unpaired two-tailed t-test). g, Protein expression of genes related to cellular senescence. The experiment was repeated for three times. h, EdU incorporation assay. Data are presented as the mean ± s.e.m. n = 5 for each group (unpaired two-tailed t-test). i, Representative GO terms of DEGs. P values were calculated by Fisher’s exact test. j, The expression of SASPs and cell cycle-related genes. k, ChIP–qPCR using the FOXP1 antibody at the CDKN1A promoter. Data are presented as the mean ± s.e.m. n = 3 for each group (unpaired two-tailed t-test). l, Activity of WT and mutant (MT) CDKN1A promoter luciferase (luc) reporter. Empty vector-infected cells (MOCK) served as the control. Data are presented as the mean ± s.e.m. n = 6 for each group (one-way ANOVA). m, mRNA expression after FOXP1 overexpression in COV434. Data are presented as the mean ± s.e.m. n = 3 for each group (unpaired two-tailed t-test). n, SA-β-gal staining. The percentage of SA-β-gal-positive cells was shown on the right. Data are presented as the mean ± s.e.m. n = 5 for each group (unpaired two-tailed t-test). o, mRNA expression of CDKN1A. Data are presented as the mean ± s.e.m. n = 3 for each group (unpaired two-tailed t-test). p, The protein expression of CDKN1A in COV434. Data are presented as the mean ± s.e.m. This test was repeated three times (unpaired two-tailed t-test).

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