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. 2017 Nov 9;7:15173. doi: 10.1038/s41598-017-15634-9

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© The Author(s) 2017

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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Increased ANO1 expression in HPV− samples is associated with hypermethylation at positively correlated CpG’s relative to HPV+ and control tissue samples. DNA methylation in HPV+ (n = 34) and HPV− (n = 241) tumor samples as well as paired control tissue (n = 48) was compared. Using a two-way repeated measures ANOVA we found (A) a significant group effect at the DRE with Bonferroni post-hoc tests revealing hypermethylation at multiple CpG’s in HPV− tumors relative to HPV+ tumors and control tissue as well as HPV+ tumors relative to control tissue, (B) no group effect at the TSS, and (C) a significant group effect at exon 2 with Bonferroni post-hoc tests revealing hypermethylation at multiple CpG’s in HPV− tumors relative to HPV+ tumors and control tissue. Data presented as mean ± SEM. *p < 0.05, **p < 0.01, ***p < 0.001.