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. 2018 Oct 2;8:14690. doi: 10.1038/s41598-018-32927-9

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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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HDAC inhibition increases both H3K18ac and H3K18cr in a dose-dependent manner. (A) Deacetylase and (B) decrotonylase activities were measured using increasing concentrations of whole-cell extracts from mouse ES and HEK-293T cells using Boc-Lys(Ac)-AMC and BOC-Lys(Cr)-AMC substrates. Average plots of n = 3 technical replicates. ES cells were treated with either, increasing concentrations of LBH589 for 24 hrs (C), or treated with 50 nM of LBH589 for indicated time (D), before histones were extracted and subjected to quantitative western blotting using an Odyssey scanner. Levels of H3K18ac and H3K18cr were normalized to the level of histone H3 and graphs show the average normalized signal intensity (mean ± SEM; n ≥ 3). Uncropped scans of western blot gels are in Supplementary Fig. 2A,B.