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. 2017 Aug 21;7:9001. doi: 10.1038/s41598-017-07458-4

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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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Pocket wall interactions are not required for NEIL1 activity against 5caC: The glycosylase activities of NEIL1 and of the indicated variants were tested by incubation with dsDNA substrates containing a 5OHC, 5fC or 5caC base (³²P radiolabelled in the modified strand). After 1 hour incubation of 20 nM dsDNA with 200 nM enzyme at 37 °C, the samples were treated with NaOH to induce single strand breaks at abasic sites. Cleavage products were analysed by denaturing gel electrophoresis and visualized by autoradiography.