Extended Data Fig. 8: PRC2 contributes to the K36M-dependent silencing of the somatic program.
From: H3K36 methylation maintains cell identity by regulating opposing lineage programmes
(a) H3K27me3 deposition within H3K36me2 domains in K36M vs. WT samples in day-4 reprogramming intermediates. Domains gaining H3K27me3 are colored in red, domains losing H3K27me3 are colored in blue. (b) Ontology terms for genes with promoters overlapping H3K36me2 domains and gaining H3K27me3. Analysis and p values from geneontology.org. (c) Heatmaps for H3K27me3 and H3K4me3 at promoters mesenchymal and epithelial genes in WT and K36M samples. (d,e) Fraction of Epcam+ and Oct4-GFP+ cells in WT (blue) and K36M (red) samples with knockdown of indicated PRC2 components (top). Log2(fold change) of fraction normalized to control siRNA (bottom). Error bars indicate mean ± SD (n = 3 independent biological experiments). (f) Representative histograms of flow cytometry for Epcam in K36M cells with control siRNA and knockdown of Ezh2 or Suz12. (g) qRT-PCR for mesenchymal (Vim, Prrx1, Zeb1), epithelial (Cdh1, Epcam), and pluripotency (Pou5f1) marker genes, error bars indicate mean ± SD (n = 3 biologically independent experiments). (h) Immunofluorescence for H3K27me3 in WT and K36M cells transfected with control siRNA or knockdown of Ezh2 or Suz12. Representative result from three independent biological experiments.
