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. 2014 Jan 16;63(2):632–644. doi: 10.2337/db13-0981

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© 2014 by the American Diabetes Association.

Readers may use this article as long as the work is properly cited, the use is educational and not for profit, and the work is not altered. See http://creativecommons.org/licenses/by-nc-nd/3.0/ for details.

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AW-recipient mice show higher proinflammatory response in the gut mucosa. Four-week-old NOD mice were continued on AW or switched to NW for 4 weeks and killed at 8 weeks of age, and PP and intestinal tissues were used for immunological analysis. A: PP cells were activated ex vivo with phorbol myristic acid and ionomycin in the presence of brefeldin A for 4 h and stained for surface CD4 and CD8, followed by intracellular IFN-γ and IL-17 for FACS analysis. Representative FACS analysis graphs with percentage values (left panel) and mean ± SD of percentage values of IFN-γ– and IL-17–positive cells among CD4 and CD8 populations from 4 mice/group tested independently (right panel) are shown. B: cDNA prepared from the small intestine (distal ileum) was used in real-time quantitative PCR to examine the expression profiles of cytokines and transcription factors. Relative expression levels of these factors in each tissue were calculated against the expression level of a housekeeping gene (β-actin) of the same sample. Mean ± SD of values from 3 mice/group tested independently in triplicate is shown.