Journal: Nature microbiology
Article Title: IFN-I and IL-22 mediate protective effects of intestinal viral infection
doi: 10.1038/s41564-019-0470-1
Figure Lengend Snippet: Representative images of murine SI (a), murine colon (b) or human colon (c) organoids either unstimulated or stimulated with 500U/ml of IFNβ for 48 h and expression of Mx2 mRNA relative to unstimulated organoids. Scale bars represent 50μM. The following number of independent experiments were performed: (a), n=7; (b), n=3 and (c), n=3. Log 10 transformed Mx2 expression was analyzed by two-tailed paired t-test. (d) Phosphorylation of STAT3 at 2 h in SI organoids. Representative of 4 independent experiments. (e) Representative images of Reg3β expression and representative mean fluorescence intensity (MFI) of individual organoids of 2 independent experiments at 24 h. Scale bars represent 50μM. The following number of organoids were analyzed: unstimulated, n=34; IFNβ, n=32 and IL-22, n=40. ANOVA with Dunnett’s multiple comparisons test was used to analyze Reg3β expression. Proportion of IL-22 expressing ILC (f) and CD11b low CD11c + CD103 − cells (g) in ABX-treated Ifnar1 f/f and Ifnar1 ΔIEC mice at day 6 post DSS. The following number of mice were analyzed for IL-22: Ifnar1 f/f ABX, n=16; Ifnar1 f/f ABX+CR6, n=19; Ifnar1 ΔIEC ABX, n=15 and Ifnar1 ΔIEC ABX+CR6, n=15. The following number of mice were analyzed for CD11b low CD11c + CD103 − cells: Ifnar1 f/f ABX, n=18; Ifnar1 f/f ABX+CR6, n=20; Ifnar1 ΔIEC ABX, n=14 and Ifnar1 ΔIEC ABX+CR6, n=14. ANOVA with Tukey’s multiple comparisons test was used to analyze cell populations. (h) Gene expression analysis of IECs from MNV CR6-infected Ifnar1 f/f (n=3) and Ifnar1 ΔIEC (n=3) mice compared to uninfected mice ( Ifnar1 f/f , n=3 and Ifnar1 ΔIEC , n=3) at day 6 post DSS. Circles in Venn diagram represent number of transcripts that are enriched following MNV-infection in the mice with the indicated genotypes. Pathway analysis was performed on the non-overlapping gene set and p-values determined by Ingenuity Pathway Analysis. All bars represent mean and error bars represent standard error of the mean. All p-values are shown in the figure.
Article Snippet: Staining was performed using sheep anti-mouse Reg3β antibody (R&D systems) followed by detection with anti-sheep Ig NL557 (R&D systems) and mounting with Vectashield containing Dapi (Vector Laboratories).
Techniques: Expressing, Transformation Assay, Two Tailed Test, Phospho-proteomics, Fluorescence, Gene Expression, Infection
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