Fig. 6. Germline DMRs in single MII oocytes from F1 females are unaffected by dietary exposure but show an altered transcriptional programme.

a Heatmap representing mean DNA methylation levels for each gametic (g)DMR in F1^mat-CD and F1^mat-HFD oocytes (merged from 41 and 37 oocyte scBS-seq datasets, respectively). b SeqMonk screenshot showing mean DNA methylation in F1^mat-CD and F1^mat-HFD oocytes over nonoverlapping 100 CpG windows (colour-coded blocks) across a ~450 kb interval encompassing the Dlk1-Dio3 imprinted cluster with a zoomed-in region (below) showing the CpG methylation calls (methylated red; un-methylated blue) of the Dlk1-Gtl2 region with quantification over the gDMR and sDMRs. Error bars represent the standard deviation from the mean of 5 pseudo-bulk groupings of 7-8 oocytes each. c Principal component analysis of scRNA-seq datasets of individual oocytes from F1^mat-CD and F1^mat-HFD. d Heatmap revealing 5 unsupervised clusters of the 166 most variable genes between F1^mat-CD and F1^mat-HFD oocytes. Top bars identify the F1 donor and diet groups. Clusters 1 to 5 comprised 25, 62, 44, 25 and 9 genes respectively. e Major terms highlighted in the gene ontology analysis of up-regulated genes from clusters 1, 2 and 4 (x-axis, -log10 of FDR adjusted p values). Gene ontology analysis was performed with GOrilla and summarised with Revigo. f Comparison of Dlk1-Dio3 microRNA (miRs) expression in F1^mat-CD (black) and F1^mat-HFD (blue) oocytes, alongside three stably expressed miRs (27b-3p, 103-3p, 423-3p), analysed by small RNA sequencing. Each of the Dlk1-Dio3 miRs was found to be significantly more represented in F1^mat-HFD oocytes. (Bars represent mean counts per million ±SD; small RNA-seq libraries generated from oocytes from four female mice per group; unpaired two-sided t-tests with Holm-Šídák’s correction for multiple comparisons: **padj = 0.0013, ***padj = 0.0005, ****padj < 0.0001, ns=not significant). Source data are provided as a Source Data file.