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. Author manuscript; available in PMC: 2012 Apr 18.

Published in final edited form as: Clin Pharmacol Ther. 2011 Sep 28;90(5):674–684. doi: 10.1038/clpt.2011.165

Effect of myeloid zinc finger 1 (MZF-1) on the binding and activity of the MATE2-K 5′-UTR variant, g.−130G>A. (a) Electrophoretic mobility shift analysis of MZF-1, MATE2-K reference, and MATE2-K variant g.−130G>A oligonucleotides. Oligonucleotides (MZF-1: lanes 1–3 and 8; reference g.−130G: lanes 4–6; variant g.−130A: lane 7) were incubated with nuclear protein extracts (25 µg) obtained from HCT-116 (human colon carcinoma) cells. The arrow indicates the position of the DNA–protein complex. Competition reactions and supershift assay were performed using different folds (25 (lanes 2, 5) and 250 (lanes 3, 6)) of molar excess of unlabeled consensus MZF-1 oligonucleotides and antibody against MZF-1 (lane 8). (b) Luciferase activities were measured 30 h after the cotransfection of the reference or variant reporters and various amounts of MZF-1 plasmids into (i) HCT-116 and (ii) HEK-293T (human embryonic kidney) cells. The reporter activity of each construct was compared with that of the empty vector (pGL4.11b(luc2)). The data shown represent mean values ± SD from triplicate wells in a representative experiment. *P < 0.05, +P < 0.01, #P < 0.001 vs. the naive promoter activity.

Effect of myeloid zinc finger 1 (MZF-1) on the binding and activity of the MATE2-K 5′-UTR variant, g.−130G>A. (a) Electrophoretic mobility shift analysis of MZF-1, MATE2-K reference, and MATE2-K variant g.−130G>A oligonucleotides. Oligonucleotides (MZF-1: lanes 1–3 and 8; reference g.−130G: lanes 4–6; variant g.−130A: lane 7) were incubated with nuclear protein extracts (25 µg) obtained from HCT-116 (human colon carcinoma) cells. The arrow indicates the position of the DNA–protein complex. Competition reactions and supershift assay were performed using different folds (25 (lanes 2, 5) and 250 (lanes 3, 6)) of molar excess of unlabeled consensus MZF-1 oligonucleotides and antibody against MZF-1 (lane 8). (b) Luciferase activities were measured 30 h after the cotransfection of the reference or variant reporters and various amounts of MZF-1 plasmids into (i) HCT-116 and (ii) HEK-293T (human embryonic kidney) cells. The reporter activity of each construct was compared with that of the empty vector (pGL4.11b(luc2)). The data shown represent mean values ± SD from triplicate wells in a representative experiment. *P < 0.05, +P < 0.01, #P < 0.001 vs. the naive promoter activity.