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. 2019 Mar 5;8(1):1–20. doi: 10.1016/j.jcmgh.2019.02.007

Figure 1.

Figure 1

IECs up-regulate numerous ISGs in response to C parvum infection. Intestinal epithelium was harvested from ileum mucosa of neonatal piglets at the time of peak infection (days 3–5 after infection) (n = 8) and from age-matched uninfected controls (n = 4) for performance of gene expression analysis using microarrays. (A) Representative photomicrograph of ileum mucosa from an uninfected control and C parvum–infected piglet used for microarray analysis. H&E stain. Scale bar: 20 μm. (B) Villus height and crypt depth (μm) and the percentage of total villus IECs that were infected with C parvum in control (n = 4) and C parvum–infected (n = 8) piglets used for microarray analysis. Each data point represents the average of 5 measurements per piglet. Scale bars: means ± SD. **P < .01, Student t test comparison between uninfected and C parvum–infected piglets. Heat map of significantly (C) up-regulated and (D) down-regulated genes in infected (C parvum) and control (Uninf) exfoliated ileum villus epithelial cells. Each control and infected biological replicate is represented. Gene IDs:fold change are listed to the right of the heat map. Known ISGs are highlighted in red. (E) qRT-PCR analysis of porcine ileum mucosal total cellular mRNA for the presence of ISG15 mRNA. Samples were obtained from piglets used for microarray analysis at peak C parvum infection (days 3–5, n = 5) and age-matched controls (n = 5). For each sample, the Ct value for ISG15 was normalized to expression of the housekeeping gene cyclophilin (ΔCt). Fold-change differences between each sample were compared with a representative uninfected sample using the 2-ΔΔCt method. Scale bars: means ± SD. **P < .01, Student t test comparison between uninfected and C parvum–infected piglets. (F) Fluorescence in situ hybridization showing ISG15 mRNA (red fluorescence) in villus epithelial cells of ileum mucosa from piglets at peak C parvum infection and absent in villus epithelium of control piglets. Photomicrograph representative of results of 2 independent experiments. Scale bar: 50 μm.