Figure 8.

NF-κB directly binds to FBXL10 promoter. (a) PC3 or LNCaP cells treated as shown in Figure 7a were subjected to ChIP assay using a p65 antibody or IgG. (b) An NF-κB BS probe was incubated without (lane 1, negative control) or with nuclear extracts from PC3 or LNCaP cells treated as shown in a. In all, 100-fold molar excess of unlabeled oligonucleotide (cold probe) was used as competitor. The probe was replaced by the mutant probe in the last lane of each panel. (c) Mice were transfected with 2 μg pFBXL10 reporter plasmids using the hydrodynamic procedure. In addition, the mouse to the left received 1 nmol oligonucleotide decoys (details in ‘Materials and Methods section'). The mouse to the right received 1 nmol mutant oligonucleotides. Data are representative of three independent experiments. (d) PC3 cells were co-transfected with pFBXL10 or NF-κB-BS mutated pFBXL10 reporter plasmids and p65 plasmids with the weight ratio as indicated for 24 h, respectively. Renilla luciferase was used as a positive control. Results are shown as the mean (bar) ± S.D. of at least three independent experiments