Figure 5

Transcriptional factor Sp1 slightly increases BRD7 promoter activity in NPC HNE1 and 5-8F cells. Mutation of Sp1 element decreases the activity of BRD7 minimal promoter in COS7 cells. (A) 0.5 μg pGL3/-266,-212 was cotransfected with 0.25 μg SV40 β-galactosidase vector and various indicated amounts of pCMV-HA/Sp1 into HNE1 and 5-8F cells by Lipofectamine 2000 Reagent. Luciferase activity was measured in cell extracts 38 h after transfection. The SV40 β-galactosidase vector was used for normalizing transfection efficiency. Data are the means ± S.D. of three independent experiments. (B) Top: The promoter fragment (sense) sequences to construct wild type and mutant Sp1/Myc-Max binging site reporter constructs of BRD7 minimal promoter -266/-212. Bottom: 0.5 μg promoter construct pGL3/-266,-212, Sp1/Myc-Max/SM and Sp1/DM were respectively cotransfected with 0.25 μg SV40 β-galactosidase vector into COS7 or 5-8F cells by Lipofectamine 2000 Reagent. Luciferase activity was measured in cell extracts 38 h after transfection. The SV40 β-galactosidase vector was used for normalizing transfection efficiency. Data are the means ± S.D. of three independent experiments.