Extended Data Fig. 2: GC senescence in patients with ovarian aging is associated with reduced NCOA7 expression.
From: Stress granule clearance mediated by V-ATPase-interacting protein NCOA7 mitigates ovarian aging
(a) Representative KEGG pathways of DEGs in GCs between control and POI women (hypergeometric test, BH-adjusted FDR < 0.05). (b) Western blot of NCOA7 in primary hGCs from control and POI women. (c) RT‒qPCR analysis of NCOA7 mRNA in GCs between control and POI women (n = 10 biological replicates per group). ****P < 0.0001. (d) Western blot of NCOA7 in GCs between control and middle-aged participants (n = 3 biological replicates per group). (e) RT‒qPCR analysis of NCOA7 mRNA in various human embryonic tissues (22 weeks old) (n = 3 technical replicates per group). (f-g) Immunohistochemistry of NCOA7 protein in ovaries from human (f), as well as young (8-week-old) and old (10-month-old) mice (g). Red arrow: GC. Scale bar: 100 μm or 30 μm in f, 200 μm or 80 μm in g. (h) Western blot of NCOA7 protein levels in ovaries between young and old mice. (i) Immunofluorescence of NCOA7 and GM130 in primary human GCs and KGN cells. Scale bar: 20 μm or 5 μm. (j) Immunofluorescence and quantification of ROS in GCs from control, POI, and middle-aged patients (n = 20 views per group). Scale bar: 5 μm. ***P = 0.0005, **P = 0.0043. (k) CUT&Tag assay showing H3K27ac histone modification of the NCOA7 protein in GCs from control and POI women. (l) Quantification of SA-β-gal-positive cells in GCs from control, POI, and middle-aged patients in Fig. 2f (n = 5 biological replicates per group). *P = 0.0232, *P = 0.0156. (m‒n) Expression levels of p16INK4a and p21Cip1 (m) and SASP genes (n) in GCs from control, POI, and middle-aged patients (n = 4 biological replicates per group). ns P = 0.0728, **P = 0.0015, ns P = 0.0880, *P = 0.0137 in m. ns P = 0.1611, ***P = 0.0009, *P = 0.0210, ****P < 0.0001, ns P = 0.2057, ***P = 0.0003, *P = 0.0229, *P = 0.0172 in n. Data in c, e, j and l-n are expressed as means ± s.e.m. Two-tailed Student’s t-test was used in c, and One-way ANOVA with Dunnett’s post hoc test in j and l-n for group comparisons.
