Fig. 2
H3K27M reversibly induces global redistribution of H3K27me2 and H3K27me3 (a–c). G477 wild-type cell line with H3.3-K27M overexpression. a H3K27me2/3 abundance by histone mass spectrometry (n = 3 in each group, mean ± standard deviation, Student’s t-test). b Example ChIP-seq tracks of H3K27me2/3 distribution, normalized by drosophila spike-in (ChIP-Rx). c Heatmap plots of ChIP-seq signal intensity for H3K27me2 and H3K27me3 over CGIs for parental G477 (wild-type) and G477 overexpressing K27R and K27M. CGIs are separated by kmeans clustering (k = 3). d–f. BT245 with H3K27M knockout (KO) by CRISPR/Cas9. d H3K27me3 abundance by histone mass spectrometry (K27M n = 3, KO n = 6, mean ± standard deviation, Student’s t-test). Whiskers represent standard deviation. e Example ChIP-seq tracks of H3K27me2/3 distribution, ChIP-Rx normalized. f Heatmap plots of ChIP-seq signal intensity for H3K27me2 and H3K27me3 over CGIs for parental BT245 (H3K27M) and K27M knockout (K27M-KO) by CRISPR. CGIs are separated by kmeans clustering (k = 3). g H3K27me3 signal change at CGIs of BT245, K27M-KO vs. K27M, color coded for DNA methylation. y-axis shows the difference in normalized H3K27me3 levels at CGIs in K27M vs. K27M-KO (log2), while x-axis shows normalized H3K27me3 levels in non-K27M state (K27M-KO, log2). Four categories of CGIs based on H3K27me3 levels and difference are depicted by squares. h Strong correlation of H3K27me2 in H3K27M with H3K27me3 in respective isogenic non-K27M state (K27R in G477, K27M-KO in BT245). 1000 aggregate bins are ranked on x-axis based on H3K27me2 in H3K27M state (black dotted line). H3K27me3 levels in each bin for non-K27M sample are shown in blue, H3K27me2 levels in non-K27M—in red (normalized, log2). Source data are provided as a Source Data file
