Following euthanasia, mouse colons were fixed in 10% buffered formalin for 24 hours at room temperature and then embedded in paraffin. Tissues were sectioned at 5-µm thickness and stained with hematoxylin & eosin (H&E) using standard protocols. H&E stained slides were scored by two pathologists (A.N.Y. and M.A.D.). Each colon was assigned four scores based on the degree of epithelial damage and inflammatory infiltrate in the mucosa, submucosa and muscularis/serosa, as previously described [25] (link). A slight modification was made to this scoring system; each of the four scores was multiplied by 1 if the change was focal, 2 if it was patchy and 3 if it was diffuse. The 4 individual scores per colon were added, resulting in a total scoring range of 0–36 per mouse. The scores for each of five mice per treatment group (i.e., exposed to 0, 0.25, 0.5, 1.0 and 4.0% DSS) were averaged.
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