FIG. 3.

Synergistic effect of MDP or FK565 with lipid A on induction of IL-12 p35, IL-12 p40, IFN-γ, and IL-15 mRNA expression in DCs. DCs were stimulated for 6 h (or 24 h for IL-15 mRNA) at 37°C with MDP (10 μg/ml), FK565 (10 μg/ml), and lipid A (10 ng/ml). Total RNA was extracted from the cells, and reverse transcription of the RNA samples to cDNAs was performed. Real-time PCR amplification was carried out using a LightCycler-FastStart DNA Master SYBR green I kit (Roche Diagnostics). The relative induction of IL-12 p35 (A), IL-12 p40 (B), IFN-γ (C), IL-15 (D), and IL-18 (E) mRNA expression was determined after normalization using β-actin. The results are representative of three independent experiments.