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. 2019 Jun 7;10:2529. doi: 10.1038/s41467-019-10404-9

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© The Author(s) 2019

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 — Oncohistone incorporation patterns induce changes in nuclear PRC2 distribution and sterility phenotypes. a, b Live cell imaging of GFP-tagged MES-2/EZH2 (the catalytic subunit of C. elegans PRC2), and immunofluorescence of H3K27me3 and H3.3/oncohistone in pachytene nuclei of wild-type and H3.3K27M mutant (mut) (a) and H3-like K27M oncohistone (H3-like mut) (b) worms. Scale bars represent 5 µm. Chromosome X was identified by depletion of H3.3 and H3K4me3 staining shown in Supplementary Fig. 3, and is marked with an asterisk. The cartoons on the right illustrate the mutations introduced into the H3.3 protein in each strain. c Western blot of H3K27me3 and H3.3/oncohistone levels in wild-type, mut, and H3-like mut worms. All versions of H3.3 are OLLAS-tagged to distinguish them unambiguously form H3. H3 levels are shown as loading control. d Boxplots illustrating levels of fertility of wild-type (gray), mut (blue), and H3-like mut (purple) worms at 25 °C. N = 3 for all conditions. e Bar plots showing types of sterility observed in mut (blue) and H3-like mut (purple) worms at 25 °C