Figure 7

TNF-α increases AP-1-binding activity and resistin promoter activity. A. Representative EMSA showing protein binding to the AP-1 oligonucleotide in nuclear extracts of macrophages after TNF-α treatment in the presence or absence of atorvastatin or JNK inhibitor. Similar results were observed in another two independent experiments. A significant supershifted complex (super) after incubation with AP-1 antibody was observed. Cold oligo means unlabeled AP-1 oligonucleotides. B. Constructs of resistin promoter gene. Positive +1 demonstrates the initiation site for the resistin transcription. Mutant resistin promoter indicates mutation of AP-1 binding sites in the resistin promoter as indicated. C. Quantitative analysis of resistin promoter activity. Cultured macrophages were transiently transfected with pResistin-Luc by gene gun. The luciferase activity in cell lysates was measured and was normalized with renilla activity (n = 3 per group). *P < 0.001 vs. control. ⁺P < 0.01 vs. 4 h.