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. 2021 Jun 23;40(30):4930–4940. doi: 10.1038/s41388-021-01899-y

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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. 2 — A Schema of the PBRM1 promoter region with highlighting of putative E2F binding sites. B Soluble chromatin from DU-145 (left) and LNCaP-AI (right) cells was precipitated with anti-E2F1, anti-MUC1-C or a control IgG. C Soluble chromatin from DU-145 (left) and LNCaP-AI (right) cells was precipitated with anti-MUC1-C (ChIP) and then reprecipitated with anti-E2F1 or a control IgG (re-ChIP). D DU-145/tet-MUC1shRNA cells were treated with vehicle or DOX for 7 days. Soluble chromatin was precipitated with anti-E2F1 or a control IgG. The DNA samples were amplified by qPCR with primers for the PBRM1 promoter region. The results (mean ± SD of 3 determinations) are expressed as fold enrichment relative to that obtained with the IgG control (assigned a value of 1). E DU-145/CshRNA and DU-145/E2F1shRNA cells were analyzed for E2F1 and PBRM1 mRNA levels by qRT-PCR. The results (mean ± SD of 4 determinations) are expressed as relative mRNA levels compared to that obtained for CshRNA cells (assigned a value of 1). F Lysates from DU-145/CshRNA and DU-145/E2F1shRNA (left) or LNCaP-AI/CshRNA and LNCaP-AI/E2F1shRNA (right) cells were immunoblotted with antibodies against the indicated proteins.