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. 2022 Jan 13;13:185. doi: 10.1038/s41467-021-27867-4

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© The Author(s) 2022

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 — a, b Ax2 or adprt1a⁻adprt2⁻ cells were treated with phleomycin and whole cell (a) or acid (b) extracts western blotted with antibodies as indicated. Molecular weight markers are indicated in kDa. Representative pictures of three biological repeats are presented. c h3b⁻ cells containing empty vector, or expressing Flag-H3b^wt, were left untreated or exposed to phleomycin. Following preparation of denatured chromatin, Flag immunoprecipitation was performed. Input extracts or immunoprecipitates were subjected to Western blotting using the indicated antibodies. Molecular weight markers are indicated in kDa. Representative pictures of at least three independent experiments. d Sequence comparison of human histone H3.1 and H3.3 with the Dictyostelium H3 variants H3a and H3b. S10 and S28, the main ADP-ribosylation targets in vertebrates are indicated. e Ax2 or h3b⁻ cells containing empty vector, or expressing Flag-H3b^WT, Flag-H3b^S10A or Flag-H3b^S28A were left untreated or exposed to phleomycin. Following preparation of acid extracts, western blotting was performed using the indicated antibodies. ADP-ribosylated endogenous histones and Flag-H3b are highlighted. Enrichment of Flag-H3b in chromatin fractions was quantified (right panel; n = 3; individual data points are shown and error bars represent the SEM). Molecular weight markers are indicated in kDa. Source data are provided in the Source Data file.