Figure 4.

Effects of TO on NO productions, protein, and mRNA expressions in peritoneal macrophages. A) Peritoneal macrophages (3 × 10⁵ cells/well) were incubated for 1 h with rIFN-γ (10 U/mL). The peritoneal macrophages were then stimulated with TO (0.01–1 mg/mL) for 48 h. The supernatants were analyzed for NO production by Griess method as described in the method. B) Cells were pretreated with different concentrations (0.01–1 mg/mL) of TO for 12 h. Total cellular proteins (40 μg) were resolved by SDS-PAGE, transferred to nitrocellulose membranes, and detected with specific antibodies, as described in Materials and Methods. A representative immunoblot of three separate experiments is shown. C) Total RNA was prepared for real time RT-PCR analysis iNOS expressions in peritoneal macrophages. The experiment was repeated three times and similar results were obtained. Values represent means ± S.E. of three independent experiments. * p < 0.05 vs. rIFN-γ (10 U/mL); ** p < 0.01 vs. rIFN-γ (10 U/mL) significant differences between treated groups were determined by ANOVA and Dunnett’s post-hoc test.