Figure 3

Inhibition of iNOS, IL-1β and TNF-α mRNA expression by ATL in LPS-stimulated BV-2 cells. BV-2 cells were pretreated with ATL (1, 10 and 100 nM) for 30 min in the absence or presence of 100 μM Boc-2 (30 min before ATL treatment) followed by incubation with LPS (100 ng/ml). Total RNA was prepared 6 h later and expression of iNOS (A), IL-1β (C) and TNF-α (D) mRNA was measured by real-time PCR. Levels of each mRNA were normalized to those of the house-keeping gene β-actin. The expression of iNOS protein was assessed by western blot analysis 24 h later (B). Detection of β-actin was also carried out to confirm the equal loading of proteins. Each value represents the mean ± SEM for three independent experiments.*P < 0.05 compared with LPS in the absence of ATL;**P <0.01 compared with LPS in the absence of ATL; ^## P < 0.01 compared with vehicle.