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. 2010 Oct 25;6(7):655–664. doi: 10.7150/ijbs.6.655

Figure 4.

Figure 4

Co-transfection analysis of the mouse Dmrt7 promoter. a), Effects of expression of NF-Y on the transcriptional activity of the Dmrt7 5'-flanking sequence (-60/+116). Cells were transfected with 0.2µg -60/+116 construct or mutants and 0.2µg NF-Y expression plasmids (pCX-NFYa, pCX-NFYb, pCX-NFYc). b), Effects of expression of the dominant negative mutant of NF-Y on the transcriptional activity of the Dmrt7 5'-flanking sequence (-60/+116). The dominant negative NF-Ya expression plasmid NF-YAm29 (0.1µg, 0.2µg, 0.3µg) was cotransfected with 0.2µg -60/+116 construct, which inhibits Dmrt7-luciferase activity. pGL3-basic was employed as a negative control. pRT-TK served as an inner control for transfection efficiency. The relative luciferase activities to the -60/+116 construct were determined by luciferase assay. Cells were harvested 24 h post-transfection, and luciferase activity was measured and normalized to Renilla luciferase activity. All transfection experiments were repeated at least three times and the activities are showed relative to that of the -60/+116 construct, which was set to 10. The results are the mean ± S.D. of three independent experiments. *, P < 0.05; **, P < 0.001 with compared to the -60/+116 wild-type construct.