Fig. 3

Eed is required for H3K27me3 establishment and developmental gene silencing in growing oocytes. a,b Representative images (a) and quantification (b) of H3K27me3 (red) IF in Eed-wt, Eed-het, and Eed-hom SN GV oocytes. White arrowheads indicate the oocyte nucleus as defined by Lamin B1 (green). DAPI (blue) shows DNA in somatic cells. Images represent 3–4 females per genotype, with 16–21 oocytes imaged per genotype. Scale bars: 20 μm. Average intensity of Eed-wt was set to 1.0. ****P < 0.0001, Kruskal–Wallis test plus Dunn’s multiple comparisons test, error bars represent mean ± standard deviation. c Principal Component Analysis (PCA) of RNA-seq data for Eed-hom (n = 6) vs Eed-het (n = 4), Eed-wt (n = 5) and Eed-wt Cre (n = 2) controls. d–e Differential gene expression analysis of Eed-het vs Eed-hom oocytes represented by volcano plot showing logFC against statistical significance and an MDplot showing logFC against average log counts per million reads. Genes with FDR-adjusted P < 0.05 are coloured in red. Deletion of Eed resulted in 349 significant DEGs (Eed oocyte DEGs), with 343 genes upregulated and 6 genes downregulated, including Eed. f Eed transcript levels (transcripts per million reads; TPM) in Eed-wt, Eed-het and Eed-hom GV oocytes. g GO enrichment analysis of Eed oocyte DEGs representing the top 10 significantly different biological processes impacted. h Pie chart displaying the proportion of significant pathways identified using Ingenuity pathway analysis