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. Author manuscript; available in PMC: 2018 Apr 29.
Published in final edited form as: Mucosal Immunol. 2017 Aug 16;11(2):357–368. doi: 10.1038/mi.2017.55

Figure 4. Epithelial barrier integrity is impaired in the absence of IL-6.

Figure 4

8–12 week old male and female C57Bl/6 and IL-6−/− mice were treated as previously described. Experiments were performed using littermates in groups of 3 mice and repeated twice. (a) Untreated, antibiotic-treated (Abx), recolonized, and IL-6−/− mice were gavaged with 0.6 mg/kg body weight 4 kDa FITC-dextran. After four hours, sera were collected from the mice, the fluorescence at 492 nm measured, and the amount of dextran calculated. Data are the mean concentration of dextran ± SEM. Statistical analyses were performed by one-way ANOVA with Dunnet’s test. ***, P<0.001 (b) Claudin-1 (brown) in vivo was evaluated by immunohistochemistry. Representative photos from 1 of 5 IL-6+/+ and 1 of 4 IL-6−/− mice are shown at 400X. Bar=20 µm. (c) Fluorescent in situ hybridization using a universal bacterial probe (red) was performed on IL-6+/+ and IL-6−/− mice. Nuclei were labeled with bis-benzimide (blue) and the mucus layer labeled with wheat germ agglutinin (green). Representative images are shown at 400X. Bar=20 µm. (d) Measurement of the mucus layer was performed in 3 areas of each of 4–6 high-powered fields (400X) per mouse (6 IL-6+/+ and 6 IL-6−/− mice). Data are the mean ± SEM mucus thickness. An unpaired two-tailed Student’s t-test demonstrated significance. ***, P<0.001