Naive OT-II cells were differentiated and DR3 expression was analyzed by real-time PCR analysis

DR3 expression was determined relative to naive CD4 T cell expression levels and normalized to GAPDH levels in all samples. (A) Comparison of DR3 expression on CD4 cells differentiated under Th1, Th2, and Th17 conditions. (B) Time-course analysis of DR3 expression in CD4 T cells differentiating under Th17 conditions. (C) Analysis of DR3 expression in cells differentiated under different combinations of cytokines. Primers used in A–C detect all DR3 isoforms. (D) Analysis of the full-length transmembrane-containing DR3 isoform (variant 1) in Th17 and T reg cells. (E) The ratio of total transmembrane DR3 (variants 1 and 3 combined) and full-length transmembrane DR3 (variant 1) was measured in Th17 and T reg cells. Error bars represent SD.<p><b>Copyright information:</b></p><p>Taken from "TL1A–DR3 interaction regulates Th17 cell function and Th17-mediated autoimmune disease"</p><p></p><p>The Journal of Experimental Medicine 2008;205(5):1049-1062.</p><p>Published online 12 May 2008</p><p>PMCID:PMC2373838.</p><p></p

(A) The addition of TL1A during naive T cell activation increases IL-2 production

Naive CD4 T cells were stimulated with plate-bound anti-CD3/CD28 antibody under the indicated conditions. Culture supernatants were analyzed for IL-2 24 h later by ELISA. Data represent the mean of triplicate cultures, and error bars represent SD. Results are representative of three independent experiments with similar results. (B) TL1A regulates Th17 differentiation. Naive CD4 T cells were stimulated by plate-bound anti-CD3/CD28, in the presence of control Ig or 1 μg/ml Fc-TL1A, and under Th17-differentiating conditions in the absence or presence of anti–IL-2 antibodies. The percentages of IL-17– and IFN-γ–producing cells were measured by intracellular cytokine staining. Data are a representative of three independent experiments with similar results.<p><b>Copyright information:</b></p><p>Taken from "TL1A–DR3 interaction regulates Th17 cell function and Th17-mediated autoimmune disease"</p><p></p><p>The Journal of Experimental Medicine 2008;205(5):1049-1062.</p><p>Published online 12 May 2008</p><p>PMCID:PMC2373838.</p><p></p

(A) In vitro–differentiated Th1 and Th17 cells were left unstimulated or restimulated on day 5 with WT or TL1A KO BMDCs for 24 h

Proliferation was measured by [H]thymidine incorporation during the last 8 h of stimulation. (B) In vitro–differentiated Th17 cells were stimulated with WT or TL1A KO BMDCs with and without Fc-TL1A for 24 h. Proliferation was measured by [H]thymidine incorporation during the last 8 h of stimulation. (C) In vitro–differentiated Th1 and Th17 cells were stimulated with WT or TL1A KO BMDCs pulsed with a titrating dose of OT-II peptide. The proliferation of Th1 and Th17 cells was determined at 24 h of stimulation by [H]thymidine incorporation. The horizontal dashed line represents the baseline proliferation of Th1 or Th17 cells alone. Data are representative of three independent experiments. Error bars represent SD.<p><b>Copyright information:</b></p><p>Taken from "TL1A–DR3 interaction regulates Th17 cell function and Th17-mediated autoimmune disease"</p><p></p><p>The Journal of Experimental Medicine 2008;205(5):1049-1062.</p><p>Published online 12 May 2008</p><p>PMCID:PMC2373838.</p><p></p

Naive OT-II CD4 T cells were stimulated with either WT or TL1A KO BMDCs under polarized Th1- and Th17-differentiating conditions

Cells were harvested on day 6 and restimulated with PMA plus ionomycin, followed by intracellular cytokine staining for IFN-γ and IL-17. Dot plots represent the IFN-γ and IL-17 expression among CD4 cells (percentages are shown). Data are representative of three independent experiments.<p><b>Copyright information:</b></p><p>Taken from "TL1A–DR3 interaction regulates Th17 cell function and Th17-mediated autoimmune disease"</p><p></p><p>The Journal of Experimental Medicine 2008;205(5):1049-1062.</p><p>Published online 12 May 2008</p><p>PMCID:PMC2373838.</p><p></p

(A) Restriction map of the mouse locus and the thymidine kinase (TK)– and neomycin (neo)-containing targeting construct derived from it

Restriction enzyme sites indicated are as follows: Ba, BamHI; Bg, BglII; E, EcoRI; and X, XbaI. Exons are represented as black boxes, and arrows indicate the direction of transcription. (B) RT-PCR analysis of TL1A mRNA in TL1A KO and WT kidneys. (C) Surface phenotype of naive WT (white bars) and TL1A KO (gray bars) lymph node cells. (left) Percentages of total lymphocytes positive for the indicated markers are plotted. (right) Percentages of CD4 or CD8 T cells positive for the indicated marker combinations are plotted. Data represent means of eight animals per group, and error bars are SEM.<p><b>Copyright information:</b></p><p>Taken from "TL1A–DR3 interaction regulates Th17 cell function and Th17-mediated autoimmune disease"</p><p></p><p>The Journal of Experimental Medicine 2008;205(5):1049-1062.</p><p>Published online 12 May 2008</p><p>PMCID:PMC2373838.</p><p></p

WT and TL1A KO mice were primed with MOG-CFA, and draining lymph nodes and spleen were harvested on day 10

Single-cell suspensions were stimulated with PMA plus ionomycin with Golgi Plug for 4 h, stained, and analyzed by flow cytometry. Percentages of IL-17– and/or IFN-γ–secreting cells among CD4 cells are shown. Values shown are means ± SEM ( = 12 mice per group). Results are compiled from two experiments with similar results. Differences between WT and KO mice are significant for each of the cytokine-expressing populations (IL-17, P = 0.004; IFN-γ, P = 0.002; IL-17IFN-γ, P = 0.019; IFN-γIL-17, P = 0.011; and IL-17IFN-γ, P = 0.005, as determined by the Student's test).<p><b>Copyright information:</b></p><p>Taken from "TL1A–DR3 interaction regulates Th17 cell function and Th17-mediated autoimmune disease"</p><p></p><p>The Journal of Experimental Medicine 2008;205(5):1049-1062.</p><p>Published online 12 May 2008</p><p>PMCID:PMC2373838.</p><p></p

(A and B) In vitro–differentiated Th1 and Th17 cells were restimulated on day 5 after activation with the indicated doses of anti-CD3 with or without 1 μg/ml of exogenous Fc-TL1A or control human Ig

Proliferation of Th1 cells (A) and Th17 cells (B) was determined by [H]thymidine incorporation after 24 h of stimulation. In a parallel experiment, culture supernatant was analyzed for IFN-γ (A) and IL-17 (B) expression by ELISA. Data are representative of three independent experiments. (C) In vitro–differentiated Th1 and Th17 cells were restimulated with the indicated doses of exogenous Fc-TL1A. (left) The proliferation of Th1 and Th17 cells was determined by [H]thymidine incorporation after 24 h of stimulation. (right) The proliferation of Th1 and Th17 cells in the absence or presence of 1 μg/ml Fc-TL1A was represented. Results are representative of three independent experiments. (D) TL1A specifically enhances Th17 cell proliferation. In vitro–differentiated Th1 and Th17 cells, and n–T reg and i–T reg cells were stimulated with control Ig or 1 μg/ml Fc-TL1A for 24 h. Proliferation was determined by [H]thymidine incorporation during the last 8 h of stimulation. Proliferation in the presence of positive controls was as follows: anti-CD3 for Th1 and Th17 cells (14,000 and 30,000 CPM, respectively), and IL-2 for n–T reg and i–T reg cells (98,000 and 25,000 CPM, respectively). Data are representative of two independent experiments with similar results. Error bars represent SD.<p><b>Copyright information:</b></p><p>Taken from "TL1A–DR3 interaction regulates Th17 cell function and Th17-mediated autoimmune disease"</p><p></p><p>The Journal of Experimental Medicine 2008;205(5):1049-1062.</p><p>Published online 12 May 2008</p><p>PMCID:PMC2373838.</p><p></p

Characterization of cGAS enzyme activity.

<p>Measurement of cGAMP production was conducted by LC-MS as described in Methods. (A) Time course of cGAS (15 nM) activity; (B) titration of dsDNA activation of cGAS (1nM) activity; (C) cGAS enzyme titration; (D) inhibition of cGAS (1 nM) activity by CuBr.</p

Binding affinities and <i>in vitro</i> activities of cGAS inhibitors.

<p>Binding affinities and <i>in vitro</i> activities of cGAS inhibitors.</p

Characterization of cGAMP FP assay.

<p>(A) mAb titration with Cy5-cGAMP (2 nM); (B) competition of Cy5-cGAMP (2 nM) binding to mAb 80–2 with: cGAMP, cAMP, cGMP, ATP or GTP; (C) Z’ results of FP assay in subset screen; (D) Distribution of compound activity from subset screen.</p