Fig. 5

RIF1 regulates MuERV-L interaction with H3K4me3 and H3k9me3. A Various concentrations of mCherry-Trim21 mRNA and RIF1 antibody were evaluated with the developmental abilities of embryos. Top: Various concentrations of mCherry-Trim21 mRNA. Bottom: Various concentrations of RIF1 antibody. B The remaining protein intensities of endogenous RIF1 were obtained between the RIF1 KD and control groups when the 10 ng/μL RIF1 antibody was electroporated into the oocytes. Bottom: Statistical results are shown. C The expression of MuERV-L in the 2-cell embryos of the RIF1 KD and control groups. Top: Representative immunofluorescence (IF) images of 2-cell embryos. Bottom: Statistical results from IF and qPCR. D IF images of 5mC and 5hmC enrichment during ZGA between the RIF1 KD and control groups. The statistical diagrams and the ELISA results of 5mC are shown at the bottom. E IF images of H3K4me3 and H3K9me3 enrichment during ZGA between the RIF1 KD and control groups. F ChIP-seq track signals of H3K4me3, H3K9me3 and H3K27me3 from mouse oocytes to blastocysts, which were extracted from GSE73952 and GSE 98149, and visualized by Integrative Genomics Viewer (IGV). G Track signals of H3K4me3, H3K9me3, H3K27me3, 5mC, and mRNA expression at the MuERV-L-int locus in control and RIF1 KD ESCs, which were extracted from GSE98356. Throughout, n=20 in each group for IF analysis, and n = 3 in each group for qPCR analysis. Data are presented as means ± SD. P values are calculated by Student’s t test. *p < 0.05, **p < 0.01, ***p < 0.001. Scale bars are shown in the lower right corner of the captures. RIF1 KD: RIF1 knockdown. ♀ represents maternal pronuclear (PN). ♂ represents paternal PN