Fig. 2

SFB mono-colonization causes CD-like inflammation in Tnf ^ΔARE mice. Germ-free Tnf ^ΔARE mice and littermate WT controls were monocolonized with SFB from birth. A Severity of inflammation was assessed by evaluating the tissue pathology score in tissue sections of distal ileum, cecum, and proximal and distal colon from 12-week-old WT mice as well as 4-week-old and 12-week-old Tnf ^ΔARE mice. B Quantitative analysis of SFB abundance in intestinal contents from distal ileum, cecum, and proximal and distal colon of 12-week-old WT and Tnf ^ΔARE mice. C FISH by using the eubacterial probe EUB338 was performed on tissue sections from distal ileum and colon of 12-week-old Tnf ^ΔARE mice followed by immunostaining with E-Cadherin. DAPI was used for nuclear visualization. Dotted line indicates thickness of inner mucus layer. D H&E-stained tissue sections of 12-week-old Tnf ^ΔARE mice revealed profound intestinal inflammation in the compartments of distal ileum, cecum, proximal colon, but not jejunum and distal colon when compared to 12-week-old WT controls, but also 4-week-old Tnf ^ΔARE mice. Scale bar represents 200 or 100 μm as indicated. E Quantitative analysis of Tnf and Il-17A transcript levels in mucosal tissue of distal ileum, cecum, and proximal colon from 4 and 12-week-old Tnf ^ΔARE mice and 12-week-old WT controls. Statistical significance was assessed by One-way analysis of variance (ANOVA) followed by Tukey test. *p < 0.05; **p < 0.01; ***p < 0.001 was considered statistically significant. F Quantitative analysis of Muc2, Defa5, Ang4, and Reg3b transcript levels in mucosal tissue of distal ileum from 4- and 12-week-old Tnf ^ΔARE mice and 12-week-old WT controls. Statistical significance was assessed by One-way analysis of variance (ANOVA) followed by Tukey test. *p < 0.05; **p < 0.01; ***p < 0.001 was considered statistically significant. G Immunofluorescence (IF) co-staining of Lysozyme (red) and E-cadherin (IEC borders, blue) counterstained with Dapi (nuclei, cyan) in ileal tissue sections from 4 weeks and 12 weeks mono-colonized WT and Tnf ^ΔARE mice (× 600) lower panel: higher magnification of the indicated sections (× 3600). H Quantification of the total number of Lysozyme positive Paneth cells per crypt based on IF staining. Statistical analyses were performed by one-way analysis of variance (ANOVA) followed by Tukey test