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. 2018 Oct 25;8:15791. doi: 10.1038/s41598-018-34000-x

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© The Author(s) 2018

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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MacUDG exhibits glycosylase activity specific for uracil while MacUDG-like is inactive. 5′-Cy5-labeled ssDNA (A: lanes 2–4, 8–11; B: lanes 2–4, 8–10) or dsDNA (A: lanes 5–7, 12–15; B: lanes 5–7, 11–16) were incubated without (A: lanes 2, 5, 8, 12; B lanes 2, 5, 8, 11, 14) or with 1 μM MacUDG-like (A: lanes 3, 6, 9, 13; B: lanes 3, 6, 9, 12, 15), 1 nM MacUDG (A): lanes 10, 14), and 1 μM MacUDG (A: lanes 4, 7, 11, 15; B: lanes 4, 7, 10, 13,16) at 37 °C for 10 min. The products were treated with alkaline and heating, which cleaved DNA at AP site. DNA substrates are indicated at the top of the panels; dA, normal ssDNA; dA/dT, normal dsDNA; dU, dI, and dX, damaged ssDNA; dU/G, dI/T, and dX/C, damaged dsDNA; dG/dT, mismatched dsDNA. Asterisks represent Cy5-labeled strands. Cleavage products were separated by 8 M urea-10% PAGE. M, DNA marker (A,B, lane 1). The cropped gels are used in the figure, and the full-length gels are presented in Supplementary Fig. S11.