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ajt4283-sup-0001-suppmatS1.pdf175KFigure S1: Frequency of lymph node CD4+ and CD8+ T cells in the lymph nodes of nonmanipulated B6 Lepob/ob and control mice. The cells from the draining lymph nodes were obtained, stained with anti-CD4 (A) and anti-CD8 (B) monoclonal antibodies and analyzed by flow cytometry (n = 5 mice/group). Data are representative of two independent experiments. The results are shown as the mean ± SEM. No statistical differences were observed among the groups.
ajt4283-sup-0002-suppmatS2.pdf124KFigure S2: IL-17 does not influence fully mismatched skin allograft survival. CBA skin was transplanted onto normal or IL-17 KO B6 mice. Signs of allograft rejection were monitored daily, and survival is reported for individual animals as the time of graft rejection (n = 4 animals/group). The data are representative of two independent experiments.
ajt4283-sup-0003-suppmatS3.pdf42KFigure S3: Tregs and Th17 cells from Lepob/ob naïve T cells are more effectively induced than those from WT naïve T cells. Naïve (CD4+CD62L+CD44CD25) T cells from wild-type or Lepob/ob B6 mice were cultured for 5 days in the presence or absence of TGF-β, IL-6 and leptin. Treg and Th17 differentiation of naïve CD4+ T cells from WT and Lepob/ob mice. The induction of Foxp3+ CD4+ T cells and IL-17+ CD4+ T cells was evaluated through the intracellular staining of Foxp3 and IL-17, respectively. The data are representative of three independent experiments. Lepob/ob: CD4+ T cells from Lepob/ob cultured in the absence of leptin; Lepob/obLep: CD4+ T cells from Lepob/ob cultured in the presence of recombinant leptin; WT: CD4+ T cells from wild-type mice. Data are represented from three independent experiments. The results are shown as the mean ± SEM. **p < 0.01.
ajt4283-sup-0004-suppmatS4.pdf140KFigure S4: CD4+ T cell frequency in RAG−/- skin-transplanted mice. A total of 5×.106 purified CD4+ T cells from wild-type, Lepob/ob or Lepdb/db mice were transferred to skin-grafted RAG−/− mice. At the time of the last graft rejection, the lymph nodes were obtained and stained with an anti-CD4 antibody. The cells were analyzed by flow cytometry. The data are representative of two independent experiments. (n = 3 mice/group).

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