(A) mice were treated with DSS in the drinking water for 6 d, followed by water for 10 d

This protocol was repeated for a total of three cycles. After the last cycle, cells isolated from the colon were restimulated in vitro with PMA/ionomycin for 5 h and subjected to intracellular staining for GFP, IL-17, and Foxp3. Histograms (from left to right) report percent GFPTCR-β cell subsets in total T cells (see ), total numbers of RORγt Tαβ cells present in the organ, and the ratio of IL-17–producing to Foxp3 cells within RORγt Tαβ cells (see ). Right panels show immunofluorescence histology of a colon from a healthy or a treated mouse. Bar, 100 μm. (B) mice were infected intranasally with 100 PFUs of influenza A virus for 7 d. Cells were then isolated from the lung and processed as in A. Right panels show immunofluorescence histology of a lung from healthy or an infected mouse. Bar, 50 μm. (C) Cells were isolated from the mesenteric LNs of a 4-mo-old × mouse and processed as in A. Right panels show immunofluorescence histology of a mesenteric LN from a normal or a tumor-bearing mouse. Bar, 100 μm. Data shown are representative of at least three independent experiments. Three to four mice were analyzed per group. *, P < 0.05 as compared with control (mock-treated or WT mice).<p><b>Copyright information:</b></p><p>Taken from "In vivo equilibrium of proinflammatory IL-17 and regulatory IL-10 Foxp3 RORγt T cells"</p><p></p><p>The Journal of Experimental Medicine 2008;205(6):1381-1393.</p><p>Published online 9 Jun 2008</p><p>PMCID:PMC2413035.</p><p></p

MACS-sorted naive (CD62L) CD4 T cells from the spleens of mice were stimulated in duplicates with anti-CD3 and anti-CD28 in the presence of blocking anti–IFN-γ and anti–IL-4 antibodies and the indicated cytokines or RA

After different periods of time, cells were restimulated with PMA/ionomycin for 5 h and analyzed by flow cytometry for the expression of GFP, Foxp3, IL-17, and IL-10. All plots are gated on TCR-β cells, except plots for IL-10 that are gated on GFPTCR-β cells. Numbers indicate percent cells in quadrants. Data are representative of three independent experiments.<p><b>Copyright information:</b></p><p>Taken from "In vivo equilibrium of proinflammatory IL-17 and regulatory IL-10 Foxp3 RORγt T cells"</p><p></p><p>The Journal of Experimental Medicine 2008;205(6):1381-1393.</p><p>Published online 9 Jun 2008</p><p>PMCID:PMC2413035.</p><p></p

(A and B) Flow cytometry analysis of cells isolated from the organs of 8–12-wk-old mice

Plots are gated on CD3 cells (A) or GFP CD3 cells (B). Numbers indicate mean percent cells in quadrants ± SD obtained with at least three mice. LPLs, lamina propria lymphocytes isolated from small intestine; mLN, mesenteric LNs; BM, bone marrow. (C) Immunofluorescence histology of RORγt cells in the small intestine of mice. Most RORγt cells in villi are T cells, whereas RORγt cells in cryptopatches located between crypts are CD3 LTi cells. Bar, 50 μm. (D) Expression of CD4 and CD8α by spleen GFPTCR-β and lung or GFPTCR-δ cells.<p><b>Copyright information:</b></p><p>Taken from "In vivo equilibrium of proinflammatory IL-17 and regulatory IL-10 Foxp3 RORγt T cells"</p><p></p><p>The Journal of Experimental Medicine 2008;205(6):1381-1393.</p><p>Published online 9 Jun 2008</p><p>PMCID:PMC2413035.</p><p></p

(A) Cells isolated from the spleen and mesenteric LNs of mice were sorted into eight distinct populations based on their expression of GFP, CD3, TCR-β, TCR-δ, CD4, and CD25 (Fig

S1), and gene expression was assessed using real-time PCR. Ct values were normalized to the mean Ct of five housekeeping genes. Data are the mean of two or three independent experiments. (B) Foxp3 RORγt T cells express IL-10. Cells isolated from LNs of mice were restimulated in vitro with PMA/ionomycin for 5 h and subjected to intracellular staining for GFP, IL-17, Foxp3, and IL-10 or an isotype control. Numbers indicate percent cells in quadrants. Results are representative of at least three individual experiments.<p><b>Copyright information:</b></p><p>Taken from "In vivo equilibrium of proinflammatory IL-17 and regulatory IL-10 Foxp3 RORγt T cells"</p><p></p><p>The Journal of Experimental Medicine 2008;205(6):1381-1393.</p><p>Published online 9 Jun 2008</p><p>PMCID:PMC2413035.</p><p></p