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. 2022 Sep 7;144(5):1027–1048. doi: 10.1007/s00401-022-02489-2

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© The Author(s) 2022

Open AccessThis 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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 3 — Reduced interaction with DNA methyltransferases leads to global hypomethylation in H3.3G34R-mutant pHGG. A Bubble plot showing differential enrichment of DNMT1 and DNMT3A with each oncohistone relative to WT control. Size = log2(mutant/WT). Color is a function of -log10 p-value and direction of interaction change. Statistically significant (p < 0.05) differential interactions have a black border. B Proximity ligation assays in MO3.13 cells between HA-H3 and DNMT1. Scale bar: 20 μm. Results are representative of two biological replicates and show mean foci counts relative to H3.3WT ± standard error. n: EV = 52; H3.3WT = 73; H3.3G34R = 61. p: ANOVA. C Median methylation beta value per sample from pHGG categorized as WT (n = 98); mutant for K27M (n = 34), G34R (n = 24), or IDH (n = 25); or normal brain (n = 7). The box marks the interquartile range (IQR) and shows the median value. The whiskers extend to 1.5 × IQR. p: ANOVA. D Median methylation beta value per probe from pHGG categorized as WT (n = 98); mutant for K27M (n = 34), G34R (n = 24), or IDH (n = 25); or normal brain (n = 7). The box marks the interquartile range (IQR) and shows the median value. The whiskers extend to 1.5 × IQR. p: ANOVA. E Scatter plot of number of differentially methylated regions (DMR) detected in H3.3G34R-mutant (n = 24) vs WT (n = 98) tumors assigned to each genomic location category versus the log2-enrichment against an expected background. F Enrichment of DMRs detected in H3.3G34R-mutant (n = 24) vs WT (n = 98) tumors around transcription start sites (TSS). G Network of pathways (Reactome, KEGG, Gene Ontology Biological Processes) enriched in genes with DMRs ± 1 kb from the TSS. Node size and color reflects significance and edge thickness reflects number of genes shared between two nodes. H Peptide counts of methyl-binding domain (MBD) containing proteins identified by BioID with H3.3WT or H3.3G34R. p: t test