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. 2020 Dec 17;10:22127. doi: 10.1038/s41598-020-79126-z

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© The Author(s) 2020

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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/.

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Detailed analysis of LINE-1 promoter CpG-methylation derived from peripheral blood cellular DNA (A) and cell-free DNA (B) of 8 young individuals aged 21–29 years (left panels) and 8 elder individuals aged 50–58 years (right panels). On top, a schematic representation of a full-length LINE-1 retroelement is given showing the distribution of CpG dinucleotides (short vertical lines) and the open reading frames 1 and 2 (ORF1/2). The analyzed 436 bp region of interest is indicated by square brackets. Detailed CpG-methylation profiles of this region revealed by bisulfite genomic sequencing are shown in panels A and B. Filled circles stand for methylated CpG dinucleotides. White circles stand for unmethylated CpGs. Grey circles stand for an undefined CpG methylation status. In the panel C the result is illustrated in bar diagrams in order to visualize the most differentially methylated CpG positions. Each bar shows the frequency of methylated CpGs at the position indicated. Positions are relative to the ATG start codon of LINE-1 ORF1. An asterisk indicates the positions of the primers used for the MS-PCR shown in Fig. 1. The left diagram shows the analyses from peripheral blood cellular DNA of young (green) and elder (blue) individuals. The right diagram shows the analyses from peripheral blood cfDNA of young (green) and elder (blue) individuals.

Detailed analysis of LINE-1 promoter CpG-methylation derived from peripheral blood cellular DNA (A) and cell-free DNA (B) of 8 young individuals aged 21–29 years (left panels) and 8 elder individuals aged 50–58 years (right panels). On top, a schematic representation of a full-length LINE-1 retroelement is given showing the distribution of CpG dinucleotides (short vertical lines) and the open reading frames 1 and 2 (ORF1/2). The analyzed 436 bp region of interest is indicated by square brackets. Detailed CpG-methylation profiles of this region revealed by bisulfite genomic sequencing are shown in panels A and B. Filled circles stand for methylated CpG dinucleotides. White circles stand for unmethylated CpGs. Grey circles stand for an undefined CpG methylation status. In the panel C the result is illustrated in bar diagrams in order to visualize the most differentially methylated CpG positions. Each bar shows the frequency of methylated CpGs at the position indicated. Positions are relative to the ATG start codon of LINE-1 ORF1. An asterisk indicates the positions of the primers used for the MS-PCR shown in Fig. 1. The left diagram shows the analyses from peripheral blood cellular DNA of young (green) and elder (blue) individuals. The right diagram shows the analyses from peripheral blood cfDNA of young (green) and elder (blue) individuals.