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. 2020 Feb 10;11:809. doi: 10.1038/s41467-020-14595-4

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© The Author(s) 2020

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. 5 — a A schematic representation of the experimental procedure to examine the effect of H3K4me on HU-treated rad53 mutants. b HU sensitivity was determined for JHD2 and jhd2Δ mutants on the RAD53 or rad53-K227A backgrounds with wild-type H3 or H3K4A mutation. c Immunoblot showing H3K4me1/2/3 levels in jhd2Δ strains. d Normalized pMET25-3HA-LRE1 expression in jhd2Δ rad53-K227A with wild-type H3 or H3K4A mutation e H3K4me3 deposition on indicated regions of pMET25-3HA-LRE1 in in jhd2Δ rad53-K227A with wild-type H3 after following the procedure show in a; all error bars represent ± SEM (n = 3 technical replicates). f 2D gel analysis of EcoRI-PstI-digested and SacI-SacI-digested fragments of the ARS305 region in jhd2Δ rad53-K227A with wild-type H3 or H3K4A mutated cells. In brief, cells were G1-released in 25 mM HU with suppressed pMET25-3HA-LRE1 transcription but with low (Off) or high (On → Off) levels of residual H3K4me. The purple arrows indicate spike signals corresponding to aberrant DNA intermediates that accumulated in HU treated rad53-K227A mutant with high level of H3K4me deposition.