FIG. 4.

Effects of recombinant human adiponectin, TNF-α, and free fatty acids on NF-κB–regulated gene expressions in HUVECs. After pretreating HUVECs with or without 100 μmol/l BAY11-7082 for 1 h, 25 μg/ml of adiponectin (A) or 10 ng/ml of TNF-α (B) were added, followed by incubation for 12 h. After HUVECs had been pretreated with or without 25 μg/ml of adiponectin for 1 h, 10 ng/ml of TNF-α or 0.4 mmol/l palmitate was added, followed by incubation for 12 h (C and D). Cell lysates from HUVECs were subjected to SDS-PAGE, followed by electrophoretic transfer to a nitrocellulose membrane. Membranes were incubated for 1 h at room temperature with antibody against ICAM-1 (A, B, and C) or FHC (D). Detection was performed using an electrochemiluminescence chemiluminescent kit according to the manufacturer's instructions. Representative data (each bracket) from four independent experiments (two samples for each experiment) are presented. Values are means ± SE. *Significant difference (P < 0.05) relative to ICAM-1 (A–C) and FHC (D) expressions in control cells in the absence of ligands. **Significant difference (P < 0.05) relative to ICAM-1 expression (C) in paired control cells in the absence of adiponectin but in the presence of palmitate (lane 3) or TNF-α (lane 5). N.S., not significant relative to FHC expression in control cells in the absence of adiponectin (A), TNF-α (B), and each ligand (D).