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. Author manuscript; available in PMC: 2013 Jul 12.
Published in final edited form as: Biochem J. 2011 Nov 15;440(1):73–84. doi: 10.1042/BJ20110392

Figure 3.

Figure 3

NF-Y binding to the Y box within the exon 1a promoter is essential for the exon 1a promoter activity. (A) Reporter gene activity was evaluated by transient transfection assays in HeLa and EA.hy926 cells. In the mutated Y box, the central CCAAT sequence was changed to CCT. (B) HeLa, EA.hy926 or THP-1 nuclear extract (NE) was incubated with a labeled probe containing the Y box, and the indicated unlabeled probes or antibodies were added for competition or super-shift analysis. The super-shifted complex is indicated by an *. (C) and (D) HeLa cells were transfected with 100 ng of the exon 1a promoter reporter plasmid along with 400 ng of NF-YA/B/C plasmids (equimolar mixtures) or 300 ng of the exon 1a promoter construct with 200 ng of dominant-negative NF-YA13m29. Luciferase activities were measured 24 hours after transfection and normalized by Renilla reporter activities. The data are shown as means and range error bars of duplicate measurements, based on the activity of +7/+266. The experiment was repeated three times with similar results.