NF-κB p65 regulates miR-17 promoter activity. (A) NF-κB p65 binding sites in the regulatory sequence of the miR-17 gene. (B) Schematic for reporter luciferase genes driven by DNA fragments in the regulatory sequence of the miR-17 gene. WT1, WT2 and WT3 represent three different binding sites. (C) Expression of reporter genes p65-BR-Luc-WT1/2/3 was regulated by p65 but not p65-BR-MUT in 293T cells. 293T cells were transfected with the indicated plasmids and incubated for 48 h. Luciferase activity was assayed after cells were collected following incubation. (D) p65 protein expression in VSMCs transfected with different amounts of p65 cDNA and Lipofectamine. (E) Expression of miR-17 in VSMCs transfected with different amounts of p65 cDNA and Lipofectamine. The relative expression levels were quantitated using densitometry. Representative results were obtained from three repeats. *P<0.05 and **P<0.01 vs. pcDNA3.1; #P<0.05 and ##P<0.01 vs. 0 µg p65 cDNA/0 µl Lipofectamine. NF-κB, nuclear factor-κB; miR, microRNA; WT, wild-type; MUT, mutant; VSMCs, vascular smooth muscle cells.