FIG. 2.

Effects of IκB/NF-κB inhibitor on FHC in murine primary cultured skeletal muscle cells. Primary cultured skeletal muscle cells were pretreated with 100 μmol/l BAY11-7082 or 50 μg/ml NF-κB SN50 at 1 h before the cells were incubated with 40 μg/ml of adiponectin for 12 h. Cells were lysed in ice-cold lysis buffer and centrifuged at 14,000g for 10 min at 4°C. Supernatants including tissue protein extracts were subjected to SDS-PAGE. Transferred membranes were incubated for 1 h at room temperature with antibody against phosphor–IκB-α (A, upper panel), IκB-α (A, lower panel), p65 NF-κB (B), and FHC (C and D). After blotting with the indicated secondary antibody, detection was performed using an electrochemiluminescence chemiluminescent kit. Representative data (one sample each for A and B; two each for C and D) from four independent experiments (two samples for each experiment) are presented. Values are means ± SE. *Significant difference (P < 0.05) relative to IκB-α phosphorylation (A, upper panel), total IκB-α (A, lower panel), p65 NF-κB (B), or FHC expression (C and D) of control cells in the absence of adiponectin. **Significant difference (P < 0.05) relative to IκB-α phosphorylation (A, upper panel) or total IκB-α (A, lower panel) of paired control cells in the absence of adiponectin (lane 5). N.S., not significant relative to IκB-α phosphorylation (A, upper panel), total IκB-α (A, lower panel), p65 NF-κB (B), or FHC expression (C and D) of paired control cells in the absence of adiponectin.