Figure 6

Western blot analysis to determine type of activation of adult primary microglial cells produced from wild-type and IL-1 KO mice stimulated with IL-4, IL-13 or IL-4/IL-13 with or without IL-1β. (A) Representative western blotting data of primary microglial cells produced from wild-type (wild) and IL-1 KO mice and exposed for 24 hours to IL-4, IL-13 or IL-4 plus IL-13 (IL-4/IL-13) with or without IL-1β. Each lane expected to CD206 blotting were applied 7 μg of reduced samples. Non-reduced samples (5 μg) were applied to detect CD206. Densitometric analysis of COX2 (B), Ym1 (C), Arg-1 (D) and CD206 (E) (n = 3 each group). (B) COX2 levels are increased by exposure of cells to IL-1β and are not influenced by IL-4 or IL-13 alone. The COX2 level was slightly enhanced by IL-1β and IL-4 co-treatment. (C) Ym1 levels are increased by exposure of cells to IL-4 and IL-4/Il-13 and are synergistically increased by co-treatment with IL-1β. However, only a low level of Ym1 is seen upon exposure of cells to IL-13, and is significantly less than that seen in response to exposure of cells to IL-4. (D) Arg-1 shows similar levels in response to exposure to IL-4 and IL-4/IL-13; these are synergistically increased by co-treatment with IL-1β. However, low levels of Arg-1 are seen for exposure of cells to IL-13. (G) CD206 was detected in response to exposure of cells to both IL-4 and IL-4/IL-13 with or without IL-1β; however, CD206 levels in IL-13-exposed samples were lower than those seen with the other treatments. Data are expressed as mean ± SD (n = 3). *: P < 0.05, **: P < 0.01, ***: P < 0.001 compared with the IL-4-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; COX 2, cyclooxygenase 2.