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. 2021 Jul 6;12:4147. doi: 10.1038/s41467-021-24380-6

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© The Author(s) 2021

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. 1 — a Sequence alignment of ERG domain containing the EZH2 recognized R-K-S motif from diverse species. b Detection of methylated ERG in VCaP cells by immunoblotting with anti-mERG antibody and competition with methylated (M) and non-methylated (C) peptides (n = 2). c Detection of mERG in VCaP cells by immunofluorescence microscopy with anti-mERG antibody pre-incubated with the specific competitor and control peptides (n = 2). Scale bar = 20 µm. d Detection of ERG, EZH2, and mERG in VCaP cells by immunofluorescence microscopy (n = 2). Scale bar = 20 µm. e Detection of ERG and mERG by IB in control and HA-tagged K362A ERG transfected VCaP cells (n = 2). f In vitro methylation assay with recombinant ERG and EZH2 followed by immunoblots with indicated antibodies (left) and in the presence of the EZH2 inhibitor GSK343 (right) (n = 2). g Detection of mERG, ERG, and EZH2 by IB in VCaP cells upon EZH2 knockdown by two siRNA (siEZH2 and siEZH2 3′UTR) (n = 2). h Detection of mERG, ERG, and EZH2 by IB in VCaP cells upon treatment with 10 µM DZNep (H) at indicated time points (n = 2). i Immunoblots of mERG, ERG, and EZH2 in RWPE1 cells transiently transfected with the indicated ERG and EZH2 expression vectors (n = 2). j Binding of recombinant ERG and EZH2 determined by microscale thermophoresis (MST). Insert, MST tracing. k Co-IP of ERG and EZH2 in PC3 cells transiently transfected with Ha-tagged ERG expression vector (n = 2). l Co-IP of ERG and EZH2 in VCaP cells with ERG and EZH2 specific antibodies and control IgG (n = 2). m Diagram of truncated ERG constructs. n Co-IP and His-pulldown in PC3 cells transiently transfected with the His-ΔN-ERG constructs and immunoblotting with anti-His and anti-EZH2 antibodies (n = 2). o Diagram of truncated EZH2 constructs. p Binding of Myc-EZH2-∆SET to Ha-ERG assessed by co-immunoprecipitation in PC3 cells transiently transfected with the truncated EZH2 constructs along with ERG plasmid (n = 2). Molecular weights are indicated in kilodaltons (kDa). Source data are provided as a Source Data File.