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. 2020 Jan 17;11:338. doi: 10.1038/s41467-019-14219-6

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© The Author(s) 2020

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. 3 — a Soluble chromatin from wild-type (WT) LNCaP and LNCaP-AI cells was precipitated with anti-AR or a control IgG. The DNA samples were amplified by qPCR with primers for the BRN2 promoter. The results (mean±SD of three determinations) are expressed as the relative-fold enrichment compared to that obtained with the IgG control (assigned a value of 1). b Schema of the BRN2 promoter region with positioning of the putative MYC binding motifs. c Soluble chromatin from LNCaP-AI cells was precipitated with anti-MUC1-C, anti-MYC or a control IgG (left). Soluble chromatin from LNCaP-AI cells was precipitated with anti-MUC1-C (ChIP) and then reprecipitated with anti-MYC or a control IgG (re-ChIP) (right). d LNCaP-AI/tet-MUC1shRNA cells were treated with vehicle or 500 ng/ml DOX for 7 days. Soluble chromatin was precipitated with anti-MYC or a control IgG. The DNA samples were amplified by qPCR with primers for the BRN2 promoter. The results (mean ± SD of three determinations) are expressed as the relative-fold enrichment compared to that obtained with the IgG control (assigned a value of 1). e LNCaP-AI cells were transfected with pGL3-Basic Luc, pBRN2-Luc (WT), pBRN2-Luc MUT1 or pBRN2-Luc MUT2 for 48 h and then analyzed for luciferase activity. The results (mean±SD of three determinations) are expressed as relative luciferase activity as compared to that obtained for cells transfected with the pBRN2-Luc (WT) vector (assigned a value of 1). f LNCaP-AI/tet-CshRNA (left panel), LNCaP-AI/tet-MUC1shRNA (middle panel) and LNCaP-AI/tet-MYCshRNA (right panel) cells treated with vehicle or 500 ng/ml DOX for 5 days were transfected with pGL3-Basic Luc or pBRN2-Luc vectors for 48 h and then analyzed for luciferase activity. The results (mean±SD of three determinations) are expressed as relative luciferase activity as compared to that obtained for untreated cells (assigned a value of 1). g LNCaP-AI cells expressing a tet-CshRNA or tet-MYCshRNA were treated with vehicle or 500 ng/ml DOX for 5 days. Lysates were immunoblotted with antibodies against the indicated proteins. *p < 0.05 (Student’s t-test). Source data are provided as a Source Data file.