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. Author manuscript; available in PMC: 2014 Sep 19.
Published in final edited form as: Nat Immunol. 2009 Mar 8;10(4):427–436. doi: 10.1038/ni.1717

Figure 6.

Figure 6

CD27 signals regulate the differentiation of γδ thymocytes. (a) Quantitative RT-PCR analysis of the expression of Ifng and Tnfa in γδ27+ cells sorted from TCRα-deficient mice (a) and TCRβ-deficient mice (b), presented as described in Figure 2a. Data are representative of two independent experiments (error bars, s.d.; n=3 mice). (b) Gene expression and fluorescence intensity of microarray analysis of wild-type and CD27- deficient (KO) γδ T cells. Black, genes with differences in expression of over twofold (WT/KO); gray, all other genes analyzed. P<0.00003 (adjusted) for genes selected as having differences in expression. Data are representative of two independent experiments. (c) Semiquantitative RT-PCR analysis of the expression of various genes (left margin) in total γδ thymocytes from wild-type or CD27-deficient mice. Data are representative of two independent experiments with triplicate samples of cells purified from four to six mice. (d) Quantitative RT-PCR analysis of the expression of Ltbr in total γδ thymocytes sorted from wild-type and CD27-deficient mice, presented as described in Figure 2a. *, P<0.001 (Student's t-test). Data are representative of two independent experiments (n=3 mice). (e) Semiquantitative RT-PCR analysis of the expression of various genes (left margin) in γδ subsets from wild-type thymi. Data are representative of two independent experiments with triplicate samples of cells purified from four to six mice. (f) Quantitative RT-PCR analysis of the expression of Ifng and II17 in γδ T cell subsets sorted at day 7 from FTOCs treated with a soluble recombinant fusion protein of CD70 and immunoglobulin (10 mg/ml; sCD70 +) or control human immunoglobulin G (sCD70 -), presented as described in Figure 2a. Data are representative of two independent experiments (error bars, s.d.; n=3 mice). *, P<0.05 (Student's t-test). (g) Flow cytometry of FTOCs at day 7, treated with medium alone, anti-CD3ε (0.67 mg/ml), anti–keyhole limpet hemocyanin (0.67 mg/ml; isotype control) or cyclosporin A (0.5 mM). Data are representative of two independent experiments with a pool of three E14 thymic lobes each.