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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. 5 — A Schema of the SLC7A11 promoter region with highlighting of the NRF2 binding site in intron-1. Soluble chromatin from DU-145 cells was precipitated with anti-NRF2, anti-MUC1-C, anti-PBRM1 or a control IgG (left). Soluble chromatin from DU-145 cells was precipitated with anti-NRF2 (ChIP) and then reprecipitated with anti-MUC1-C, anti-PBRM1 or a control IgG (re-ChIP). B Soluble chromatin from DU-145/tet-MUC1shRNA cells treated with vehicle or DOX for 7 days was precipitated with anti-NRF2 (left), anti-PBRM1 (right) or a control IgG. The DNA samples were amplified by qPCR with primers for the SLC7A11 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). C Chromatin from DU-145/tet-MUC1shRNA cells treated with vehicle or DOX for 7 d was analyzed for ATAC-seq. UCSC genome browser snapshot of ATAC-seq data from the SLC7A11 gene showing loss of peaks and decrease in chromatin accessibility as a function of MUC1-C silencing. D and E. DU-145/tet-MUC1shRNA cells treated with vehicle or DOX for 7 days (D) and DU-145/CshRNA and DU-145/PBRM1shRNA cells (E) were analyzed for SLC7A11 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)(left). Lysates were immunoblotted with antibodies against the indicated proteins (right). F Schema of the G6PD promoter region with highlighting of the NRF2 binding site in intron-2. Soluble chromatin from DU-145 cells was precipitated with anti-NRF2, anti-MUC1-C, anti-PBRM1 or a control IgG (left). Soluble chromatin from DU-145 cells was precipitated with anti-NRF2 (ChIP) and then reprecipitated with anti-MUC1-C, anti-PBRM1 or a control IgG (re-ChIP)(right). The DNA samples were amplified by qPCR with primers for the G6PD 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). G Chromatin from DU-145/tet-MUC1shRNA cells treated with vehicle or DOX for 7 d was analyzed for ATAC-seq. UCSC genome browser snapshot of ATAC-seq data from the G6PD gene showing loss of peaks and decrease in chromatin accessibility as a function of MUC1-C silencing. H and I. DU-145/tet-MUC1shRNA cells treated with vehicle or DOX for 7 days (H) and DU-145/CshRNA and DU-145/PBRM1shRNA cells (I) were analyzed for G6PD 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) (left). Lysates were immunoblotted with antibodies against the indicated proteins (right).