Figure 7

Expression of H3.3K4M mutant impaired H3R2me2s accumulation. (a), (c) Immunofluorescent images of H3K4me3 (red) or H3R2me2s (red) and DAPI (blue) in H3.3-EGFP-expressed, H3.3R2A-EGFP-expressed, H3.3K4M-EGFP-expressed, and untreated zygotes. EGFP (green) shows successful injection. Key: ♂, male pronuclei; ♀, female pronuclei. Scale bar = 20 µm. (b), (d) Quantification of the H3K4me3 and H3R2me2s signal intensities in pronuclei. Red bars indicate the mean values. The mean value of male pronuclei in untreated zygotes was set as 1. One-way ANOVA and the Tukey–Kramer test were performed to evaluate statistical significance. *Significantly different from the control (**p < 0.01, ***p < 0.001); n.s.: Not significantly different from the control. The number of zygotes in (b) was 15 for the H3.3-EGFP expressed group, 10 for the H3.3R2A-EGFP expressed group, and 13 for the untreated group. The pronucleus used for quantification are presented in Fig. S11a. The number of zygotes in (d) was 22 for the H3.3-EGFP expressed group, 29 for the H3.3K4M-EGFP-expressed group, and 23 for the untreated group. The pronucleus used for quantification are presented in Fig. S13.