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. 2022 Oct 27;50(19):10929–10946. doi: 10.1093/nar/gkac861

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© The Author(s) 2022. Published by Oxford University Press on behalf of Nucleic Acids Research.

This is an Open Access article distributed under the terms of the Creative Commons Attribution-NonCommercial License (https://creativecommons.org/licenses/by-nc/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact journals.permissions@oup.com

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Figure 1. — EZH2 non-canonically binds genomic sites with gene-active markers, Pol II and AR or AR-V7, in prostate cancer cells. (A, B) Averaged intensities (A) and heatmaps (B) of EZH2 or H3K27me3 CUT&RUN and ATAC-seq signals ± 5 kb from the centers of canonical EZH2⁺/H3K27me3⁺/low-accessibility peaks (i.e. EZH2-ensemble, 8436 peaks; top panels) or non-canonical EZH2⁺/H3K27me3⁻/high-accessibility peaks (i.e. EZH2-solo, 15 987 peaks; bottom panels) in 22Rv1 cells. (C) Heatmap showing the signals of EZH2 (in duplicate), H3K27me3 (in duplicate), H3K27ac (in triplicate), H3K4me2, H3K4me3, BRD4 and RNA Pol II ± 5 kb from the centers of EZH2-solo (top) or EZH2:PRC2-ensemble peaks (bottom). Except H3K4me2, H3K4me3 and Pol II data which were from LNCaP-abl cells, all the others were generated in 22Rv1 cells. EZH2 and H3K27me3 datasets were from CUT&RUN experiments and all the others from ChIP-seq. (D, E) Heatmap showing the EZH2, H3K27me3, AR and AR-V7 signals ± 5 kb from the centers of either EZH2-solo (D) or EZH2:PRC2-ensemble (E) peaks identified in 22Rv1 cells. Mock, ligand-stripped condition; DHT, ligand-treated condition. (F) Venn diagram showing the overlapping of EZH2-solo peaks with either AR (left) or AR-V7 peaks (right) in 22Rv1 cells. (G) Venn diagram showing a significant overlap between EZH2-solo:AR- and EZH2-solo:AR-V7-co-occupied sites in 22Rv1 cells. (H) Pie chart showing genomic annotation of EZH2-solo:AR (left) or EZH2-solo:AR-V7 (right) peaks in 22Rv1 cells. (I, J) Averaged intensities (top) and heatmaps (bottom) of EZH2, H3K27me3 and AR ChIP-seq signals ± 5 kb from the centers of those EZH2-solo:AR-co-occupied sites identified from 22Rv1 cells, in LNCaP-abl (I) or VCaP (J) cells. (K) Heatmap showing the signals of AR (only 22Rv1 cells shown; left) or EZH2 binding, ± 5 kb from the centers of those EZH2-solo:AR-co-bound peaks identified in 22Rv1 cells (second column), in the AR-negative DU145 and PC3 cells (two right columns).