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. 2022 May 25;20(9):1379–1390. doi: 10.1158/1541-7786.MCR-22-0165

Figure 3.

Figure 3. MUC1-C activates NOTCH2 expression by a MYC-mediated mechanism. A, Schema of the NOTCH2 promoter and enhancer regions with locations of potential E-boxes. B and C. Soluble chromatin from H69 cells was precipitated with anti-MUC1-C, anti-MYC, or a control IgG (B). Soluble chromatin was precipitated with anti-MUC1-C (ChIP) and then reprecipitated with anti-MYC or a control IgG (re-ChIP; C). The DNA samples were amplified by qPCR with primers for the NOTCH2 promoter and enhancer regions. D, Soluble chromatin from H69/tet-MUC1shRNA cells treated with vehicle or DOX for 7 days was precipitated with anti-MYC or a control IgG. The DNA samples were amplified by qPCR with primers for the NOTCH2 promoter region. The results (mean ± SD of three determinations) are expressed as the relative fold enrichment compared with that obtained with the IgG control (assigned a value of 1). E, H69/tet-MUC1shRNA (left) and DMS53/tet-MUC1shRNA (right) cells treated with vehicle or DOX for 7 days were analyzed for NOTCH2 mRNA levels by qRT-PCR. F, H69/tet-MYCshRNA (left) and DMS53/tet-MYCshRNA (right) cells treated with vehicle or DOX for 7 days were analyzed for NOTCH2 mRNA levels by qRT-PCR. G, COR-L279/tet-MUC1shRNA (left) and H526/tet-MUC1shRNA (right) cells treated with vehicle or DOX for 7 days were analyzed for NOTCH2 mRNA levels by qRT-PCR. The results (mean ± SD of four determinations) are expressed as relative mRNA levels compared with that obtained for vehicle-treated cells (assigned a value of 1).

MUC1-C activates NOTCH2 expression by a MYC-mediated mechanism. A, Schema of the NOTCH2 promoter and enhancer regions with locations of potential E-boxes. B and C. Soluble chromatin from H69 cells was precipitated with anti-MUC1-C, anti-MYC, or a control IgG (B). Soluble chromatin was precipitated with anti-MUC1-C (ChIP) and then reprecipitated with anti-MYC or a control IgG (re-ChIP; C). The DNA samples were amplified by qPCR with primers for the NOTCH2 promoter and enhancer regions. D, Soluble chromatin from H69/tet-MUC1shRNA cells treated with vehicle or DOX for 7 days was precipitated with anti-MYC or a control IgG. The DNA samples were amplified by qPCR with primers for the NOTCH2 promoter region. The results (mean ± SD of three determinations) are expressed as the relative fold enrichment compared with that obtained with the IgG control (assigned a value of 1). E, H69/tet-MUC1shRNA (left) and DMS53/tet-MUC1shRNA (right) cells treated with vehicle or DOX for 7 days were analyzed for NOTCH2 mRNA levels by qRT-PCR. F, H69/tet-MYCshRNA (left) and DMS53/tet-MYCshRNA (right) cells treated with vehicle or DOX for 7 days were analyzed for NOTCH2 mRNA levels by qRT-PCR. G, COR-L279/tet-MUC1shRNA (left) and H526/tet-MUC1shRNA (right) cells treated with vehicle or DOX for 7 days were analyzed for NOTCH2 mRNA levels by qRT-PCR. The results (mean ± SD of four determinations) are expressed as relative mRNA levels compared with that obtained for vehicle-treated cells (assigned a value of 1).