Figure 2 from MUC1-C Dictates PBRM1-Mediated Chronic Induction of Interferon Signaling, DNA Damage Resistance, and Immunosuppression in Triple-Negative Breast Cancer
Year: 2025
Type: other
Abstract: MUC1-C regulates PBRM1 expression and function in activating the STAT1 and IRF1 genes. A, Genome browser snapshots of ATAC-seq data from the PBRM1 gene in BT-549/tet-MUC1shRNA cells treated with vehicle or DOX for 7 days (left). Chromatin was analyzed for accessibility by nuclease digestion (right). The results are expressed as the percentage of undigested chromatin (mean ± SD and individual values). B, BT-549/tet-MUC1shRNA cells treated with vehicle or DOX for the indicated days were analyzed for PBRM1 mRNA levels by qRT-PCR (left). The results (mean ± SD and individual values) are expressed as relative mRNA levels as compared with that obtained in control vehicle-treated cells (assigned a value of 1). Lysates were immunoblotted with antibodies against the indicated proteins (right). C, Schema of the PBRM1 gene with localization of a PLS downstream to the TSS. Soluble chromatin from BT-549/tet-MUC1shRNA cells treated with vehicle or DOX for 7 days was precipitated with a control IgG, anti–MUC1-C and anti-IRF1. The DNA samples were amplified by qPCR with primers for the PBRM1 PLS region. The results (mean ± SD and individual values) are expressed as relative fold enrichment as compared with that obtained with IgG (assigned a value of 1). D, BT-549/CshRNA, BT-549/IRF1shRNA and BT-549/IRF1shRNA#2 cells were analyzed for PBRM1 mRNA levels by qRT-PCR (left). The results (mean ± SD and individual values) are expressed as relative mRNA levels as compared with that obtained in control vehicle-treated cells (assigned a value of 1). Lysates were immunoblotted with antibodies against the indicated proteins (right). E, Nuclear lysates from BT-549 cells were precipitated with a control IgG and anti–MUC1-C. Input proteins and precipitates were immunoblotted with antibodies against the indicated proteins. F, Soluble chromatin from BT-549/tet-MUC1shRNA cells treated with vehicle or DOX for 7 days was precipitated with a control IgG and anti-PBRM1. The DNA samples were amplified by qPCR with primers for the indicated IRF1 and STAT1 regions. The results (mean ± SD and individual values) are expressed as fold-enrichment as compared with that obtained from control IgG-precipitated chromatin (assigned a value of 1). G, Genome browser snapshots of ATAC-seq data from the indicated IRF1 regions in BT-549/CshRNA and BT-549/PBRM1shRNA cells. Chromatin from the indicated IRF1 regions was analyzed for accessibility by nuclease digestion. The results are expressed as the percentage of undigested chromatin (mean ± SD and individual values). H, Genome browser snapshots of ATAC-seq data from the STAT1 PLS in BT-549/CshRNA and BT-549/PBRM1shRNA cells. Chromatin was analyzed for accessibility by nuclease digestion. The results are expressed as the percentage of undigested chromatin (mean ± SD and individual values). *, P ≤ 0.05; **, P ≤ 0.01; ***, P ≤ 0.001; ****, P ≤ 0.0001.
Authors: Nami Yamashita, Yoshihiro Morimoto, Atsushi Fushimi, Rehan Ahmad, Atrayee Bhattacharya, Tatsuaki Daimon, Naoki Haratake, Yuka Inoue, Satoshi Ishikawa, Masaaki Yamamoto, Tsuyoshi Hata, Sayuri Akiyoshi, Qiang Hu, Tao Liu, Henry Withers, Song Liu, Geoffrey I. Shapiro, Tomoharu Yoshizumi, Mark D. Long, Donald Kufe
Language: English
Subfield: Immunology
Field: Immunology and Microbiology
Domain: Life Sciences
Open Access status: gold
OpenAlex ID: W4416764786