Fig 2: Ovarian senescence is increased in doxorubicin-treated mice. a Schematic diagram for animal experiment design. b–d Body weight, ovary weight, and ovary index of mice after saline or doxorubicin injection (n = 16, unpaired Student’s t-test). e The proportion of regular or irregular estrous cycles after saline or doxorubicin injection (n = 16, Chi-square test). f The serum levels of AMH, FSH, and E2 at diestrus (n = 6, unpaired Student’s t-test). g The proportion of follicles at different stages (PMF: primordial follicles, GF: growing follicles, ATF: atretic follicles). h Representative H&E staining images of murine ovaries. Yellow arrows: primordial follicles, red arrows: primary follicles, blue arrows: secondary follicles, black arrows: zona pellucida remnants. ANF: antral follicles. i Follicle counting results based on H&E staining of serial ovarian sections (n ≥ 5, unpaired Student’s t-test). j Representative images of SA-β-gal staining and IHC detection of p16 and p21 in the murine ovaries. k IHC scores of relative expression of p16 and p21 based on IOD (n = 3, unpaired Student’s t-test). IOD: integrated optical density. l, m Protein expression of p16 and p21 in ovarian tissues detected by western blot. n Relative mRNA expression of genes related to cellular senescence and SASP (n = 3, unpaired Student’s t-test). Data are presented as mean ± SD. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001

Fig 3: Senolytics can reduce ovarian senescence in doxorubicin-treated mice. a Representative SA-β-gal staining images of murine ovarian sections. b, c Representative images of p16 and p21 expression in murine ovaries by IHC. d, e IHC scores of relative expression of p16 and p21 based on IOD (n = 3, one-way ANOVA). f, g Protein expression of p16 and p21 in ovarian tissues shown by western blot. h Relative mRNA expression levels of genes related to cellular senescence and SASP in ovaries (n = 3, one-way ANOVA). Data are presented as mean ± SD. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001