plasticine
Figure 2: Dendritic cells expressing FasL (FasL-DCs) reduce interferon-γ production and induce apoptosis in vitro. A. Western blot to determine the expression of FasL by bone marrow-derived dendritic cells transfected with 5, 10, or 15 μg of FasL-coding vector or 15 μg of empty vector (EV-DCs). Membrane-bound (44 KDa) and soluble (26 KDa) forms of FasL were observed. B. ELISPOT to determine IFN-γ production by EV-DCs and FasL-DCs cultured alone with or without treatment with formyl-methionyl-leucylphenylalanine (fMLP) or cocultured with CD4+ T cells and treated with fMLP. n=3 per group. *** p<0.001 vs negative control, ###p<0.001 vs EV-DCs and CD4+ T cell coculture. C. Flow cytometry for annexin-V staining (AU) in cocultures of T cells and EV-DCs or FasL-DCs that were treated with fMLP. Representative flow cytometry blots for panel C are shown in Supplemental Figure 2.