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. 2020 Mar 20;10:5181. doi: 10.1038/s41598-020-62035-6

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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 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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Fgf21 mRNA expression in Hepa1-6 cells with targeted DNA demethylation of the Fgf21 promoter induced using the dCas9-SunTag and scFv-TET1CD system. (a) Experimental protocol of the transient transfection system with gRNAs (scramble gRNA, gRNA1, gRNA2, and gRNA1+2) and administration of SPPARMα, K-877 (left panel). Fgf21 mRNA expression levels after the administration of K-877 (right panel). n = 7–9 per group. (b) Experimental protocol of the stable transfection system with scramble gRNA, gRNA 1, and gRNA 1+2 and administration of K-877 (upper panel). Fgf21 mRNA expression levels after the administration of K-877 (lower left panel). n = 6–7 per group. Correlation between Fgf21 mRNA levels post K-877 administration and %DNA methylation (n = 12) (lower right panel). Statistical analyses were performed using Spearman’s rank correlation coefficient. CI: Confidence interval. (c) ChIP assays of Pol II, histone markers, Dnmts, and PPARα. Primers amplifying the regions −106 to +21 (for Pol II, histone markers, and Dnmts) and of −977 to −923 bp (for PPARα) were used for ChIP-qPCR analysis (Fig. 1b). n = 5–6 per group. Statistical analyses were performed using the Mann–Whitney U-test. Data are expressed as mean ± SD. *P < 0.05, **P < 0.01; ***P < 0.001; ****P < 0.0001; N.S., not significant between the denoted pairs.