Figure 3. Expression of N1 and N2 receptors regulate Th17 and RORγt ⁺FOXP3⁺; FOXP3⁺ T_reg cell frequencies in the gut.

Lymphocytes were isolated from the LP of small intestine (SI) and colon of naïve N1N2^lox/lox and N1N2^CD4Cre mice. IL-17A frequency in CD45⁺ CD4⁺ CD3⁺ cells was assessed by flow cytometry on PMA/Ionomycin stimulated cells in the SI and colon. (a) Numbers in representative flow cytometry plots as well as histograms on the right show the mean frequency of IL-17A⁺ in gated CD45⁺ CD4⁺ CD3⁺ T cells in the SI and colon. (b) Representative flow cytometry plots and histrogram bars of the frequency of IL-17A⁺ RORγt⁺ and IL-17A⁻ RORγt⁺ cells gated on CD45⁺ CD4⁺ CD3⁺ T cells in the SI and colon (c) Representative flow cytometry and histogram of the mean frequency of RORγt ⁺FOXP3⁺ in CD45⁺ CD4⁺ CD3⁺ T_reg cells present in the SI is shown. (d) Representative flow cytometry plots and histrograms show the mean frequency of FOXP3⁺GATA3⁺ in CD45⁺ CD4⁺ CD3⁺ T_reg cells in the SI. Mean values ± SEM (n ≥ 3 per group) are shown. Data are shown as a pool of ≥3 mice per group; *p < 0.05, **p < 0.01 versus control, unpaired, two-tailed student’s test.