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. 2018 Jun 5;8:8637. doi: 10.1038/s41598-018-26882-8

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© The Author(s) 2018

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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NNMT overexpression reduces the liver SAM/SAH ratio and decreases the level of CTGF promoter methylation. (A) SAM levels, SAH levels, and SAM/SAH ratios in the livers of NMMT Tg mice (black bars) and WT littermates (white bars) on an HFD with NAM. The data represent the means ± SEM (n = 3 animals/group). (B) Relative GNMT, GAMT, PEMT and BHMT1 mRNA expression in the liver. (n = 3 per group). (C) Bisulfite sequencing of the CTGF, Col4a1 and Col4a2 CpG island. Each circle indicates a CpG site in the sequence, and each line of circles represents analysis of a single cloned allele. ○, unmethylated CpG sites; •, methylated CpG sites. Abbreviations: NNMT, nicotinamide N-methyltransferase; SAM, S-adenosylmethionine; SAH, S-adenosylhomocysteine; HFD, high fat diet; NAM, nicotinamide; Tg, transgenic; WT, wild type; GNMT, glycine N-methyltransferase; GAMT, guanidinoacetate N-methyltransferase; PEMT, phosphatidylethanolamine N-methyltransferase; BHMT, betaine homocysteine methyltransferase; MSP, methylation-specific polymerase chain reaction; CTGF, connective tissue growth factor; COL, collagen.