Extended Data Fig. 3: NCOA7 deficiency accelerates cellular senescence and ovarian aging.
From: Stress granule clearance mediated by V-ATPase-interacting protein NCOA7 mitigates ovarian aging
(a) Quantification of the apoptosis rate in WT and NCOA7-KO KGN cells (n = 5 biological replicates per group). ns P = 0.3981. (b) Quantification of cell proliferation in WT, NCOA7-KO, and NCOA7-rescued KGN cells (n = 5 biological replicates per group). Scale bar: 5 μm. The rescued group was compared with the KO group (#). ####P < 0.0001, ****P < 0.0001, ####P < 0.0001, ****P < 0.0001. (c) Quantification of IL-1β, IL-8, MMP3 and CCL2 in the supernatants of WT, NCOA7-KO, and NCOA7-rescued KGN cells (n = 5 biological replicates per group). The rescue group was compared with the KO group (#). *P = 0.0172, #P = 0.0206, *P = 0.0115, #P = 0.0279, **P = 0.0030, ##P = 0.0054, **P = 0.0025, #P = 0.0181. (d) Western blot of p16INK4a and p21Cip1 in WT, NCOA7-KO, and NCOA7-rescued KGN cells. (e) PCR genotyping of WT and Ncoa7−/− mice. (f) Western blot of ovarian tissues to confirm absence of NCOA7 protein expression in Ncoa7−/− mice. (g) The serum levels of E2 and FSH in 10-month-old WT and Ncoa7−/− mice (n = 5 biological replicates per group). *P = 0.0224, *P = 0.0412. (h) The ovary index in 10-month-old WT and Ncoa7−/− mice (n = 4 biological replicates per group). ns P = 0.6741. (i) Immunohistochemistry analysis of p16INK4a and p21Cip1 in ovaries of WT and Ncoa7−/− mice (n = 3 biological replicates per group). Scale bar: 100 μm. *P = 0.0135, ***P = 0.0006. (j-k) Quantification of cell proliferation (n = 3 biological replicates per group) (j) and expression levels of SASP genes determined via RT‒qPCR (n = 4 biological replicates per group) (k) in GCs from WT and Ncoa7−/− mice. ***P = 0.0010, ***P = 0.0003 in (r). ns P = 0.0661, **P = 0.0072, *P = 0.0241, *P = 0.0196 in (s). (l) Quantification of E2 production in the supernatants of GCs isolated from WT and Ncoa7−/− mice (n = 4 biological replicates per group). *P = 0.0106. Data in a-c, g-h, and j-l are expressed as means ± s.e.m, and compared by two-tailed Student’s t-test.
