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. 2021 May 26;11:11023. doi: 10.1038/s41598-021-89389-9

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© The Author(s) 2021

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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Characterization of the PTEN promoter in A375 and A375PR1 cell lines. (a) qRTPCR measuring PTEN promoter CpG methylation, as determined by the remaining (unmethylated) DNA upon treatment with the methyl-cytosine-specific restriction enzyme McrBc (n = 3). (b–c) ChIP followed by qRTPCR analysis measuring the levels of (b) EZH2 and (c) H3K27me3 at the PTEN promoter in A375 and A375PR1 cell lines (n = 3). (d–e) qRTPCR measuring baseline expression levels of (d) DNMT3A and (e) EZH2 in A375 and A375PR1 cell lines (n = 3). (f–g) qRTPCR measuring the expression levels of PTEN following siRNA induced knockdown of (f) DNMT3A and (g) EZH2 in A375 and A375PR1 cell lines (n > 3). (h) qRTPCR measuring the expression levels of PTEN following simultaneous siRNA-induced knockdown of EZH2 and DNMT3A in A375 and A375PR1 cell lines (n = 3). (a–h; p-values represent a two-tailed student’s t-test).