FIGURE 3.

Time-dependent activation of Egr-1, NFκB, and AP-1 DNA binding and activity in response to globular adiponectin. A, RAW 264.7 macrophages were treated with 1 μg/ml gAcrp for 0–18 h. Nuclear extracts were then prepared and used to measure the binding of nuclear proteins to oligonucleotides specific for Egr-1, NFκB, AP-1, and SP-1 DNA binding sites. Images are representative of at least three independent experiments. B, RAW 264.7 macrophages were co-transected with either pAP1-Luc or NFκB-Luc cis reporter plasmids. Twenty-four hours after transfection, cells were then treated with 1 μg/ml gAcrp for 0–18 h, and luciferase mRNA were measured by quantitative real time PCR. Values are expressed relative to β-actin mRNA (means ± S.E., n = 4–5). *, p < 0.05 compared with cells not treated with gAcrp.