Figure 8.

Effect of p38 inhibitor SKF86002 on p38 activities, TNF-α production, and TNF-α promoter activity. (A) MC/9 cells sensitized with anti-OVA IgE were incubated in the presence of 10 μg/ml OVA for 5 min and p38 activities were measured. SKF 86002 at 1–10 μM significantly inhibited p38 activity. The data are expressed as the percentage of kinase activity detected in the presence of 10 μg/ml OVA and 0.1% DMSO (mean ± SD, n = 4; ∗∗, P < 0.01). (B) MC/9 cells sensitized with anti-OVA IgE (1 × 10⁶ per ml) were incubated with 0.1% DMSO or 10 μM SKF 86002 for 2 h. The cells were then incubated with 10 μg/ml OVA for 3 h. SKF 86002 at 10 μM did not affect TNF-α production (mean ± SD, n = 4; NS, not significant). (C) pTNF(-1311)Luc (4 μg) was transfected into MC/9 cells. MC/9 cells were passively sensitized with anti-OVA IgE after the transfection and were incubated for an additional 15 h with 10 μg/ml OVA or PBS in the presence of 0.1% DMSO (0) or 10 μM SKF 86002. Luciferase activities were measured as relative light units (RLU) and standardized by control RLU (PBS) (mean ± SD, n = 4; NS, not significant). (D) pCMV5ΔMEKK1 (MEKK1, 2 μg) or pCMV5 empty plasmid (pCMV5, 2 μg) was transfected into MC/9 cells with pTNF(-1311)Luc (2 μg) in the presence of 0.1% DMSO (0) or 10 μM SKF 86002. Luciferase activities were measured as relative light units 24 h after the transfection (mean ± SD, n = 4; NS, not significant).