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. 2019 Jun 20;9:8853. doi: 10.1038/s41598-019-45356-z

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© The Author(s) 2019

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 methylcytosine content delays ExoIII-mediated resection; but does not affect the rate of resection of either ExoIII or T7 Exonuclease. (a) 5 nM and (b) 10 nM ExoIII was added to a non-methylated substrate, a substrate containing one methylated cytosine, and a substrate containing four methylated cytosines (*S, _*S^1M and _*S^4M, respectively). Standard curve is represented by the grey dotted lines. (c) Time (seconds) until the ExoIII reaction reaches completion on the methylated and unmethylated substrates based on the point at which the graphs plateau in (a) and (b). (d) Calculated resection rate of ExoIII based on maximum gradients in (a) and (b). (e) 6 nM and (f) 12 nM T7 Exo on non-methylated and differentially methylated substrates. Standard curve is represented by the grey dotted lines. (g) Time (seconds) until the T7 Exo reaction reaches completion on the methylated and unmethylated substrates based on point at which the graphs plateau in (e) and (f). (h) Calculated resection rate of ExoIII. Error bars represent SEM; n = 3 in all cases; *p < 0.05, **p < 0.01, ***p < 0.001.