Figure 3.

Aciculatin suppresses the increase of mRNA and protein expression and promoter activity of LPS-induced iNOS and COX-2 in macrophages. A. 1 × 10⁶ RAW264.7 cells were treated with aciculatin (1-10 μM) for 30 min, then stimulated with LPS (1 μg/mL) for 5 h, mRNA of iNOS and COX-2 were measured by RT-PCR. B. Treatment of RAW264.7 macrophages with aciculatin (1-10 μM) for 30 min followed by stimulation with LPS (1 μg/mL) for 24 h in the continued presence of aciculatin. Then the cells were harvested and whole cell extracts were prepared for Western blot analysis for the indicated proteins. C. Cells (1 × 10⁵ cells) were transiently transfected with 1 μg of plasmid pGL3-miNOS or pGL3-mCOX-2 for 24 h, then were treated with 10 μM aciculatin for 30 min, followed by stimulation with LPS (1 μg/mL) in the continued presence of the aciculatin for another 24 h. Luciferase activity was then measured as described in the Materials and Methods. The results are expressed as the mean ± S.E.M. for three separate experiments, each with three replicates. * p < 0.05 and ** p < 0.01 compared with the control group; ## p < 0.01 for comparison of indicated groups.