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. 2021 Jul 28;4:919. doi: 10.1038/s42003-021-02439-w

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© The Author(s) 2021

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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Fig. 2 — Comparison between MacroGreen and the wild type Af1521-GFP fusion binding to either PARylated PARP1 (a) or MARylated PARP10 (b) coated plates. In total, 50 µl reactions in 96-well Nunc MaxiSorp^TM plates; detection with 1 µM MacroGreen or Af1521-GFP at room temperature c Examples of the linear assay range using MacroGreen detection of PARP1 PARylation predominantly at acidic side chains; PARP1 PARylation predominantly at serine side chains in the presence of HPF1; PARP10 MARylation at various side chains; or C2I MARylation of actin at Arg177. d Extended linear range of MAR detection on PARP10 under the same conditions. e Detection of MAR on five histone proteins (H2B, H3.1, H3.2, H3.3 and H4 as indicated) in the presence of PARP10. PARP10 MARylation alone (gray) serves as reference. Negative control reactions contained NAD⁺ (1 mM) but not PARP10. Statistical significance of the data was calculated using a one-way ANOVA test; error bars indicate S.D. All panels – n = 4.