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. Author manuscript; available in PMC: 2016 Apr 1.
Published in final edited form as: Oncogene. 2015 Feb 9;34(40):5187–5197. doi: 10.1038/onc.2014.442

Figure 3. MUC1-C promotes NF-κB-mediated activation of TAK1 expression.

Figure 3

A. Schematic representation of the TAK1 promoter with positioning of the NF-κB binding site (upper panel). SK-CO-1 cells were transfected with wild-type or mutant pTAK1-Luc for 48 h and then assayed for luciferase activity. The results are expressed as the relative luciferase activity (mean±SD of three determinations) compared to that obtained with wild-type pTAK1-Luc (lower panel). B. Soluble chromatin from SK-CO-1/CshRNA and SK-CO-1/MUC1shRNA cells was precipitated with anti-NF-κB p65 and, as a control, IgG. The final DNA samples were amplified by qPCR with pairs of primers for the NF-κB binding region (NBR; -326 to -317). The results (mean±SD of three determinations) are expressed as the relative fold enrichment compared to that obtained with the IgG control. C. Soluble chromatin from the indicated SK-CO-1 cells was precipitated with anti-NF-κB p65. The precipitates were released, reimmunoprecipitated with anti-MUC1-C and then analyzed for TAK1 promoter sequences. The results (mean±SD of three determinations) are expressed as the relative fold enrichment compared to that obtained with the IgG control. D. LoVo/MUC1-C cells were transfected with wild-type or mutant pTAK1-Luc for 48 h and then assayed for luciferase activity. The results are expressed as the relative luciferase activity (mean±SD of three determinations) compared to that obtained with wild-type pTAK1-Luc. E. Soluble chromatin from LoVo/vector and LoVo/MUC1-C cells was precipitated with anti-NF-κB p65 and, as a control, IgG. The final DNA samples were amplified by qPCR with pairs of primers for the NF-κB binding region (NBR). The results (mean±SD of three determinations) are expressed as the relative fold enrichment compared to that obtained with the IgG control. F. Soluble chromatin from the indicated LoVo cells was precipitated with anti-NF-κB p65. The precipitates were released, reimmunoprecipitated with anti-MUC1-C and then analyzed for TAK1 promoter sequences. The results (mean±SD of three determinations) are expressed as the relative fold enrichment compared to that obtained with the IgG control.