Fig. 7: Inflammation-induced primordial follicle loss was mediated by activation of CD38 and a decline in NAD+ levels.
a, Workflow showing LPS treatment in vivo and in vitro. b, Expression levels of Cd38 and inflammation-associated genes (Il1a, Il1b, Il6, Il10 and Tnf) by real-time RT‒PCR for ovaries 24 h after injection of LPS (n = 4 for each group). c, Western blotting analysis of CD38 and TNF protein levels in ovaries 24 h after LPS injection. d, Relative expression of each protein relative to GAPDH levels (n = 3 for each group). e, Expression of Cd38 and inflammation-associated factors in cultured ovaries from 10-day-old WT-control, LPS-treated WT, Cd38−/−-control and LPS-treated Cd38−/− mice (n = 6 for each group). f, NAD+ levels in cultured WT-control, LPS-treated WT, Cd38−/−-control and LPS-treated Cd38−/− ovaries (n = 8 experimental replicates, each using 2 or 3 ovaries). g, Representative images of ovarian sections from cultured WT-control, LPS-treated WT, Cd38−/−-control and LPS-treated Cd38−/− ovaries. The enlarged pictures show primordial follicles (arrows) and growing follicles (GFs; arrowheads) in ovaries from the different groups. The bottom images are representative of primordial follicles and GFs (primary, secondary and antral follicles) in WT mice (n = 6 ovaries for each group). Scale bar, 100 μm. h, Percentages of primordial follicles and growing follicles in ovaries for each group. i, Immunofluorescence staining of Foxo3 in ovarian sections cultured for 4 d for the different groups. Scale bar, 20 μm. j, Percentages of nuclear export of Foxo3 in primordial follicles from different groups (n = 8 ovaries for each group). Data are presented as the mean ± s.e.m. P value was determined by unpaired two-tailed t-test between the two groups. NS, not significant.
