FIGURE 1.

Stimulation with Nogo-P4 increased the expression of inflammatory mediators in primary cultured microglia. Microglia separated from the cerebral cortex were seeded into wells precoated with PBS or Rtn-P4 (100 μg/ml) or Nogo-P4 (100 μg/ml) and stimulated for 6, 12, or 24 h. Cell lysates and supernatant were collected to detect the expression of inflammatory mediators. Western blot was used to determine the expression of iNOS (A and B) and COX-2 (C and D). Values are reported as mean ± S.D. (error bars), as a percentage of values determined in the PBS group (control, 100%). The Western blot data represent the results of four independent experiments. The Griess method was performed to detect NO release (E). The proportions of PGE₂ (F), IL-1β (G), and TNF-α (H) secreted into the medium was assayed by ELISA kits. Values are reported as mean ± S.D. Griess and ELISA data represent the results of three independent experiments. Statistical analysis was performed using one-way ANOVA with post hoc Tukey's test. *, p < 0.05; **, p < 0.01; ***, p < 0.001.