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. 2017 Aug 14;8:185. doi: 10.1186/s13287-017-0638-7

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© The Author(s). 2017

Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided 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 Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.

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Fig. 3 — Nkx2.5 promoter CpG island demethylation. a Position of Nkx2.5 gene on chromosome 5, sequence of the promoter region indicated. b After bisulfite conversion, the 160-bp promoter region of Nkx2.5 was amplified and cloned for bisulfite sequencing for control (U) and differentiated cardiomyocytes (D); shaded regions indicate the CpG islands that are modified after cardiac differentiation. c CpG islands were evaluated as methylated (orange dot) and unmethylated (red dot) sites by bisulfite sequencing. d After bisulfite conversion, methylation-specific PCR was performed for Nkx2.5 using unmethylated and methylated primers in control (U) and differentiated cardiomyocytes (D) (**p < 0.01, n = 3, all experiments performed in triplicate) (Color figure online)