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Fig. 1 | BMC Biology

Fig. 1

From: Reduced PRC2 function alters male germline epigenetic programming and paternal inheritance

Fig. 1

Reduced EED function resulted in male subfertility and reduced litter size. a Average litter size produced by wild-type female mice mated to Eedwt/wt (n = 10), Eedhypo/wt (n = 13) and Eedhypo/hypo (n = 13) males mated to for two periods of 30 days. Each point represents the average of two, four or six litters per male. Average litter sizes produced by Eedhypo/hypo males were more variable in size than from Eedwt/wt and Eedhypo/wt males (Bartlett’s test P = 0.0002). b Normalised daily sperm production of Eedwt/wt (n = 3), Eedhypo/wt (n = 5), and Eedhypo/hypo (n = 6) males (mean ± SEM, one-way ANOVA; no significant differences). c Testis histology from sub-fertile and fertile Eedhypo/hypo males (top row) compared to their Eedwt/wt or Eedwt/hypo siblings (bottom images). In some Eedhypo/hypo males (top left panel), testis cords contained reduced numbers of germ cells with evidence of germ cell sloughing, while other cords demonstrated apparently normal spermatogenesis. In some Eedhypo/hypo males, testis cords appeared normal (top right panel). The total number of pups produced from the first four females is shown in parentheses. d, e Flow cytometric analysis of H3K27me3 levels (d) and cell cycle stage (e) in germ and somatic cells of E15.5 Eedwt/wt, Eedwt/hypo and Eedhypo/hypo testes. Eedwt/wt (n = 5 and 2), Eedhypo/wt (n = 7 and 4) and Eedhypo/hypo (n = 2 and 3) testes in d and e respectively. Data are mean ± SEM. f Immunofluorescent analysis of E12.5 and E14.5 Eedhypo/hypo and Eedwt/wt foetal testis sections using antibodies specific for OCT4 at E12.5 and DPPA4 at E14.5. Merged images are shown in the left panels, with greyscale images of GFP shown in the middle panels and DPPA4 in the right-hand panels. Nuclear DNA is marked by DAPI in blue. Arrows indicate germ cells that lack Oct4GFP expression. Scale bar 50 μm. g Percentage of foetuses in which Oct4GFP silencing was observed in some germ cells at E12.5 and E14.5 in Eedwt/wt (n = 0/7), Eedwt/hypo(n = 1/8) and Eedhypo/hypo (n = 6/10) animals. h Average percentage of cells in the total cell population that were Oct4GFP positive in pairs of E15.5 testes from Eedwt/wt (n = 13 from 8 litters), Eedwt/hypo (n = 31 form 17 litters) and Eedhypo/hypo (n = 14 from 9 litters) males (mean ± SEM, One-way ANOVA; No significant differences)

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