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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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Transfection of Dnmts siRNA. (a) Experimental protocol of the transient transfection system with gRNA1 and Dnmts siRNA. Closed triangles indicate BS analysis. (b) Gene expression levels of Dnmts 72 h after transfection with scramble or Dnmts siRNA in Hepa1-6 cells. Statistical analyses were performed using the Mann–Whitney U-test. Data are expressed as mean ± SD. n = 7–9 per group. **P < 0.01; ***P < 0.001; N.S., not significant vs. scramble siRNA. (c) (Left panel) BS analysis of the Fgf21 promoter at the designated time points. Representative data of four independent experiments are shown. %DNA methylation is indicated below as mean ± SD. (Right panel) Graphic presentation of statistical analysis of the BS data (n = 4 at each time point). Statistical analyses were performed using the Mann–Whitney U-test. Data are expressed as mean ± SD. *P < 0.05; N.S., not significant vs. scramble siRNA.