FIGURE 6.

Microglial activation in acute brain slices. Example of activation marker stainings on acute slices treated with LPS. (A) Immunohistochemical staining for Mac-2/Galectin-3 (red), nuclei were visualized with DAPI (blue; A1). Microglia (green) positive (A1 white square, A2) for Mac-2/Galectin-3 (red) and microglia that do not express the marker (white triangle, A3) were present in the slice. (B) Immunohistochemical staining for iNOS (red), nuclei were visualized with DAPI (blue; B1). Microglial cells that were positive (white square B1,B2) and negative (white triangle B1,B3) for iNOS (red) were observed in the slice after LPS treatment. (C) Immunohistochemical staining for IL1β (red), nuclei were visualized with DAPI (blue; C1). Microglial cells that were positive (white square C1,C2) and negative (white triangle C1,C3) for IL1β (red) were observed in the slice after LPS treatment. Examples of the different immunostainings were taken from slices treated for 24 h with 1 μg/ml LPS. Scale bar = 50 μm and for inserts = 20 μm. White squares indicate the microglia positive for the marker and shown in higher magnification (A–C2), white triangles indicate microglia negative for the marker and shown in higher magnification (A–C3). (D) Quantification of the expression of three activation markers (Mac-2, iNOS, and IL1β) by microglia in E15.5 brain slices cultured for 24 h with IL-6 (10 ng/ml), poly (I:C) (50 μg/ml), or LPS (1 μg/ml). Kruskal–Wallis test was used for statistical analysis. Number of treated slices in control and IL-6 group N = 4; LPS and poly (I:C) group N = 5. Number of cryosections for Mac-2/iNOS/IL1β in: saline group n = 23/23/27; IL-6 group n = 22/19/16; poly (I:C) group n = 18/25/19; LPS group n = 21/21/22 (all derived from three different embryos). (^∗p < 0.05).