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. 2020 Oct 30;9(11):1068. doi: 10.3390/antiox9111068

Figure 6.

Figure 6

Figure 6

IL-4 modulates MPO- and iNOS-derived oxidative/nitrosative stress within activated microglia/macrophages in the CA1 layer of pKr-2-injected hippocampus in vivo. Sections (A,D,G (PBS); B,E,H (pKr-2); C,F,I (pKr-2 + IL-4Nab)) adjacent to those used in Figure 4 were processed for immunohistochemical staining. (A–C) Double immunofluorescence images of myeloperoxidase (MPO; red) and OX-42 (green) for microglia/macrophages/neutrophils and both images are merged (yellow) in the CA1 layer of hippocampus. Scale bar, 20 μm. (D–F) Double immunofluorescence images of inducible nitric oxide synthase (iNOS; red) and OX-42 (green) for microglia/macrophages/neutrophils and both images are merged (yellow) in the CA1 layer of hippocampus. Scale bar, 20 μm. (G–I) Immunofluorescence images of nitrotyrosine (green) to detect nitrosative damages in the CA1 layer of hippocampus. Nuclei were counterstained with DAPI (blue). Scale bar, 40 μm. (J) Double immunofluorescence images of GFAP (green, left panel) for astrocytes and MPO (red, left panel) or OX-42 (green, right panel) for microglia/macrophages/neutrophils and MPO (red, both panel) and both images are merged (yellow) in the CA1 layer of hippocampus. Scale bar, 20 μm. (K) Quantification of MPO expression in OX-42+ microglia/macrophages/neutrophils. ** p < 0.001, significantly different from PBS (control). # p < 0.01, significantly different from pKr-2. mean ± SEM; n = 4 to 5 in each group, ANOVA and Newman–Keuls analysis. (L) Quantification of iNOS expression in OX-42+ microglia/macrophages/neutrophils. * p < 0.01, significantly different from PBS (control). # p < 0.01, significantly different from pKr-2. mean ± SEM; n = 4 to 5 in each group, ANOVA and Newman–Keuls analysis. (M) Quantification of nitrotyrosine expression. ** p < 0.001, significantly different from PBS (control). ## p < 0.001, significantly different from pKr-2. mean ± SEM; n = 4 to 5 in each group, ANOVA and Newman–Keuls analysis. Dotted lines indicate the CA1 layer of the hippocampus.