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. 2020 Sep 15;10:15086. doi: 10.1038/s41598-020-72155-8

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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 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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PAR levels at the regulatory regions of the MyoD target genes p57 and myogenin do not change during differentiation. (a) Western blot analysis of PARylated proteins in undifferentiated C2.7 myoblasts proliferating in growth medium (GM) and in differentiated C2.7 myotubes cultured for 24 or 48 h in differentiation medium (DM). PARylated proteins were detected with the anti-PAR antibody. ERK1 was used as an invariant control. Full length blots are presented in Fig. S10. (b) ChIP-qPCR analysis for PAR levels at KvDMR1, myogenin promoter (MyoG prom), Rhodopsin promoter (Rho prom), and Igf2 Differentially Methylated region (Igf2 DMR1) during C2.7 cell differentiation. Samples were prepared from undifferentiated cells proliferating in growth medium (GM), and from differentiated cells kept for 24 h in differentiation medium (DM) and immunoprecipitated either with PAR antibody (IP samples) or with control IgG. Protein binding is expressed as percentage of input and represents the mean ± SEM of four independent experiments for KvDMR1 and MyoG prom and of three independent experiments for Rho prom and Igf2 DMR1. Statistical significance: p-value < 0.05 (*); ns stands for not significant.