Figure 5

Western blot analysis to identify the type of activation of adult primary microglial cells from wild-type and IL-1 KO mice stimulated by vehicle, IFNγ or IL-4 with or without IL-1β. (A) Representative western blotting data of primary MG produced from wild-type (wild) and IL-1 KO mice exposed for 24 hours to vehicle, IFNγ or IL-4 with or without IL-1β. Each lane expected to CD206 blotting applied 8 μg of reduced samples. Non-reduced samples (5 μg) were applied to detect CD206. Densitometric analysis of STAT1 (B), COX2 (C), iNOS (D), Ym1 (E), Arg-1 (F) and CD206 (G) (n = 3 each group). (B) STAT1 level is increased by exposure of cells to IFNγ, but not to IL-4. The level is not influenced by co-treatment of cells with IL-1β. (C) The COX2 level is increased by exposure of cells to IL-1β but not to IFNγ or IL-4 alone. Co-treatment with IL-1β and IL-4 tends to increase the COX2 level synergistically. (D) While the iNOS level is not increased significantly by exposure of cells to IFNγ or IL-4, the level increased synergistically in response to co-treatment with IFNγ and IL-1β. (D) The level of Ym1 is increased by exposure of cells to IL-4 and further increased by co-treatment with IL-1β. (E) The Arg-1 level is also increased by exposure of cells to IL-4 and further increased by co-treatment with IL-1β. (G) The CD206 level is increased slightly by exposure of cells to IL-4, but not with co-treatment with IL-4 and IL-1β. Data are expressed as mean ± SD (n = 3). *: P < 0.05, **: P < 0.01, ***: P < 0.001 compared to the vehicle-treated group without IL-1β for each genotype (One-way ANOVA followed by Dunnett post-hoc test). ANOVA, analysis of variance; arg-1, arginase 1; COX2, cyclooxygenase 2; iNOS, inducible NO synthase.