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. 2020 Aug 6;10:13275. doi: 10.1038/s41598-020-70289-3

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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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IR induces hypermethylation of cellular proteins, but not DNA hypermethylation, in MDA-MB-231 cells. (A) Whole-cell lysates subjected to specified conditions (IR, 10 Gy) were immunoblotted with anti-asymmetrical dimethylarginine (ASYM24; full western blot) and anti-β-actin antibodies to control for equivalent loading. (B) Whole-cell lysates were immunoprecipitated with anti-Sam68 antibodies. The bound proteins were separated by SDS-polyacrylamide gel electrophoresis and immunoblotted with anti-Sam68 and ASYM24 antibodies as indicated. A control experiment in which cell lysates were immunoprecipitated with IgG is shown. (C) Effect of AA and AdOx on IR-induced methylation of BRCA1. MDA-MB-231 whole-cell lysates were immunoprecipitated with anti-BRCA1 antibody and immunoblotted with anti-ASYM24 antibody. Equal amounts of BRCA1 per lane were loaded. (D) Effect of IR (24 h), AdOx and AZA (5 µM) treatment on the genomic DNA methylation status in MDA-MB-231 cells, evaluated as the ratio of 5-methylcytosine/total cytosine (mC/tC). The histogram represents the means ± SD of three independent experiments. Statistical significance (P) was determined using Student’s paired t test, *P < 0.05 and NS (not significant) with respect to the untreated control. Except for those indicated, the groupings blots in this figure were cropped from different gels. Full blots are shown in the Supplementary Information, Fig. S6.