Fig. 4.

Separate epigenetic effects of ICSI. (a) K-means clustering for the hyper- (left) and hypo-DMRs (right) of ICSI-ET versus IVF-ET. In green-purple heatmaps, the hyper-DMRs (hypo-DMRs) were classified into seven (nine) clusters by k-means clustering on row scaled average DNA methylation level. With the same row order, row scaled DNA methylation level in each neonatal sample was also shown in cyan-red heatmaps. The number of DMRs in the selected clusters were also shown. (b) Venn diagram for the overlap between of the gain (upper) and lost (lower) H3K4me3 DPs of ICSI-ET versus CTRL, and ICSI-ET versus IVF-ET neonatal samples. (c) For the selected hyper- (union of cluster C1-C3) and hypo-DMRs (union of cluster C4-C6), gain (7233) and loss (84) H3K4me3 DPs, their gene ontology (biological process) enrichment analysis results were grouped and shown, if its enrichment terms existed (adjusted p-value < 0.05; The p-value was determined by hypergeometric test and adjusted by BH method.). (d) The chromatin states distribution of the selected hyper- and hypo-DMRs and H3K4me3 DPs (7233 GAIN, 84 LOSS) in each neonatal sample of CTRL, IVF-ET, and ICSI-ET. The chromatin states were defined in Fig. 3b.