Fig. 5

Potential regulatory effects of DNA methylation heterogeneity. a Box plots of the relationship between DNA methylation heterogeneity and expression heterogeneity in human 8-cell stage embryos. The Pearson correlation coefficient of the two heterogeneity scores was 0.18. b Volcano plot comparing the gene expression heterogeneity between mouse Stella^−/− embryos and mouse wild-type (WT) embryos at the 8-cell stage. The x-axis represents the log₂-transformed fold change, and the y-axis represents the log₁₀-transformed p value. A total of 265 genes had significantly higher expression heterogeneity scores in Stella^−/− embryos than in WT embryos, while 38 genes had significantly lower expression heterogeneity scores in Stella^−/− embryos than in WT embryos. Three genes reported to be related to cell fate determination during mouse embryogenesis are indicated in green. c Box plots comparing the DNA methylation levels in mouse zygotes between genes with increased expression heterogeneity and other genes. Student’s t test was performed (**p value < 0.01). d, e Box plots of the DNA methylation heterogeneity (d) and expression heterogeneity (e) in human 8-cell stage embryos between first cell fate determination (FCFD)-related genes and other genes. Student’s t test was performed for comparisons (***p value < 0.001). f Heatmap of the DNA methylation heterogeneity scores (blue) and expression heterogeneity scores (green) for FCFD-related genes. The FCFD-related genes were classified into four groups based on their DNA methylation and expression heterogeneity scores at the 8-cell stage (HH: DNA methylation heterogeneity > 0.3 and expression heterogeneity > 0.5; HL: DNA methylation heterogeneity > 0.3 and expression heterogeneity ≤ 0.5; LH: DNA methylation heterogeneity ≤ 0.3 and expression heterogeneity > 0.5; and LL: DNA methylation heterogeneity ≤ 0.3 and expression heterogeneity ≤ 0.5). Within the HH group, three reported ICM markers (GDF3, CUBN, and IGF1) and a TE marker (GCM1) are indicated