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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. 7 — a Immunoblots of mERG and the indicated proteins in control (EV) and stable PTEN knockdown (shPTEN) VCaP cells (n = 3). b Expression of ERG/EZH2 co-occupied genes in VCaP-shPTEN and control (EV) VCaP cells evaluated by qRT-PCR. c ERG, mERG, and EZH2 occupancy at the indicated gene promoters in VCaP-shPTEN and control (EV) VCaP cells by ChIP-qPCR. d EZH2 phosphorylation (Ps21) evaluated by IF and quantitative assessment (right) in VCaP-shPTEN and control (EV) following treatment with MK-2206 (5 µM) for 24 h. Scale bar 10 µm (n = 2). e Immunoblots of mERG and indicated proteins in VCaP-shPTEN (sh-PTEN) and control (EV) cells treated with DMSO and MK-2206 (5 µM) for 5 and 24 h (n = 2). f Diagram of WT, S21A, and S21D EZH2 mutated constructs. g Immunoblots of mERG and indicated proteins in RWPE1 cells transfected with indicated expression plasmids. Right, densitometric assessment of mERG level relative to total ERG (n = 2). h IHC evaluation of pS21 EZH2 in prostate tissue from ERG (PbCre; ERG flox/flox) and ERG/PTEN (PbCre; ERG flox/flox PTEN flox/flox) mice. Right, quantitative IHC scores of pS21EZH2 and EZH2. All error bars, mean ± s.d. (n = 3, technical replicates) (b, c). P-values were determined by one-way ANOVA test. Scale bars represent 200 µm. Source data are provided as a Source Data File.