Fig. 3: Cell cycle-dependent establishment of H3K27me3 in the male pronucleus.

a The H3K27me3 state in pronucleus at 8 h of IVF after Ezh2 mRNA microinjection. M indicates male pronucleus and F indicates female pronucleus in zygote. Scale bar, 10 μm. b Relative fluorescence intensity of H3K27me3 in PN3 stage pronucleus after Ezh2 mRNA microinjection. A1: Female pronucleus in control zygote; A2: Female pronucleus in Ezh2 mRNA microinjected zygote; A3: Male pronucleus in control zygote; A4: Male pronucleus in Ezh2 mRNA microinjected zygote. Error bars, S.E.M. P > 0.6796 and P > 0.6780 by two-tailed Student’s t tests. n.s. represents nonsignificant difference. Source data are provided as a Source data profile. c The localization of EZH2 in zygotes from PN1 to PN5 stages. Scale bar, 10 μm. d The relative level of EZH2 fluorescence intensity from PN1 to PN5 stages. n = 15 embryos at each stage examined over three independent experiments. Error bars, mean ± S.E.M. Source data are provided as a Source data profile. e The H3K27me3 state in pronucleus at 13 h of IVF after roscovitine treatment. The addition of roscovitine inhibited the establishment of H3K27me3 in the PN5 stage male pronucleus. M indicates male pronucleus and F indicates female pronucleus in zygote. Scale bar, 10 μm. f Relative fluorescence intensity of H3K27me3 in PN5 stage pronucleus after roscovitine treatment. E1: Female pronucleus in control zygote; E2: Female pronucleus in roscovitine treated zygote; E3: Male pronucleus in control zygote; E4: Male pronucleus in roscovitine treated zygote. Error bars, S.E.M. ***P < 2.05216E−17. n.s. represents a nonsignificant difference. P > 0.6963 by two-tailed Student’s t tests. Source data are provided as a Source data profile. g Western blot shows that the protein level of p-EZH2^T345 was decreased after roscovitine treatment. The median line of the box plot represents the median, and the top and bottom of the box represent the upper and lower quartile, respectively.