Figure 2

IL-17 synergizes with inflammatory cytokines to induce the expression of immune modulatory genes in MSCs. (a and c) MSCs were cultured with different combinations of inflammatory cytokines IFNγ, TNFα, and IL-17 (10 ng/ml) for 12 h, and then cells were harvested for total RNA extraction. The mRNA expression levels of iNOS (a) and chemokines CCL2, CCL5, CXCL9, CXCL10 (c) were detected by quantitative RT-PCR. (b) MSCs were cultured with IFNγ and TNFα, or together with IL-17 (10 ng/ml) for the indicated times, and the protein levels of iNOS were detected by western blotting analysis. (d) MSCs were supplemented with Sup-CD3 or Sup-CD3 pretreated with antibodies against IL-17, and cells were collected for RNA extraction after 12 h (Sup-CD3: supernatant from splenocytes activated by anti-CD3 and anti-CD28). The expression of the iNOS, CCL2, CCL5, CXCL9 and CXCL10 genes was measured by quantitative RT-PCR. (e) MSCs were treated with Sup-CD3 or Sup-CD3 pretreated with antibodies against IL-17, and the protein level of iNOS was assessed by western blotting analysis after 24 h. mRNA expression values are means±S.E.M. of three wells from a representative of three independent experiments. Western blotting data are from a representative of three independent experiments