Figure 3

Effects of SAM on TNFα-induced NF-κB activation in adipocytes. (A) After adipocytes were treated with SAM (1 mM) for the indicated days, TNFα (50 ng/ml) were treated for 18 h. Nuclear proteins were prepared and NF-κB binding activity was determined by EMSA. Lane 1 shows free probe (FP) alone. Anti-p65 antibody was incubated with the nuclear extract before the addition of the probe (lane 8). (B) After treatment of cells with TNFα or SAM, nuclear extracts and cell lysates were subjected to immunoblot analysis with anti-p65, anti-IkBα or γ-tubulin. (C) The band intensity of p65 in the nuclear extract of the cells treated with TNFα was expressed as 100 and the others were expressed as its relative values (upper panel) (n = 3). ^*P < 0.01 compared to the value of the cells treated with TNFα. ^**P < 0.05 compared to the value of the cells treated with TNFα. The band intensities of p65 in the cell lysates were normalized by those of γ-tubulin. The value of the control cells was expressed as 100 and the others were expressed as its relative values (lower panel) (n = 4). ^*P < 0.01 compared to the value of the control cells. ^**P < 0.01 compared to the value of cells treated with TNFα.