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. 2021 Sep 28;12:5618. doi: 10.1038/s41467-021-25583-7

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© The Author(s) 2021

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 2 — a Manhattan plot of the EWAS meta-analysis based on whole blood DNA methylation data from five twin cohorts (total sample size = 5723) that identified 834 MZ-DMPs. The red horizontal line denotes the epigenome-wide significance threshold (Bonferroni correction). Dark red dots highlight significant DMPs near centromeres. Orange dots highlight significant DMPs near telomeres. b Dichorionic (DC) MZ twins have separate chorions, amnions, and placentas. Monochorionic diamniotic (MCDA) MZ twins have separate amnions and a common chorion and placenta. Monochorionic monoamniotic (MCMA) have a common chorion, amnion, and placenta. It has been hypothesized that DC MZ twins result from separation soon after fertilization, whereas MC twins are thought to result from separation ≥3 days after fertilization, with MCMA twins arising later than MCDA twins. c Density plots of twin correlations for the differentially methylated positions in monozygotic twins (MZ-DMPs) identified in the EWAS meta-analysis illustrate that the overall distribution of twin correlations at MZ-DMPs show the following pattern: rMZ-MCMA > rMZ-MCDA > rMZ-DC. d Twin correlations for genome-wide autosomal methylation sites do not follow this pattern. MZ monozygotic twins, DZ dizygotic twins. e MZ-DMPs with larger correlations in monochorionic MZ twins compared to dichorionic MZ twins. The CpGs were selected by three criteria (1) rMZ-MCMA > rMZ-MCDA > rMZ-DC; (2) rMZ-MCDA > 0.5; (3) rMZ-DC < 0.2. cgid = Illumina CpG identifier. Chr = chromosome; rMZ-MCMA = correlation in monozygotic monochorionic monoamniotic pairs. rMZ-MCDA = correlation in monozygotic monochorionic diamniotic pairs. rMZ-DC = correlation in monozygotic dichorionic pairs. f–g Pathway enrichment analysis results based on the nearest genes of the 834 Bonferroni-significant MZ-DMPs identified in the meta-analysis. f Top enriched gene ontology (GO) pathways for MZ-hypo-DMPs (differentially methylated positions with a lower methylation level in monozygotic twins). The darker the color, the stronger the enrichment. g Top enriched gene ontology (GO) pathways for MZ-hyper-DMPs (differentially methylated positions with a higher methylation level in monozygotic twins). The darker the color, the stronger the enrichment.