Effects of ectopic RelB on DC maturation. Immature or CD40L stimulated DCs transduced with RelB or control vector were analyzed for DC maturation-associated molecules. (A) Overlay histograms of gated GFP⁺ cells analyzed for the indicated marker molecules. Filled histograms represent RelB-transduced cells and solid black histograms represent empty control vector transduced cells. (B) FACS diagrams of gated GFP⁺ cells analyzed for CD83 versus CD1a expression. Parallel cultures were harvested and analyzed at day 10 (top panel) or were further cultured in the presence of CD40L and then harvested 48 hours later (bottom panel). Percentages of CD1a⁺CD83⁺ cells are indicated on the plots. (C) Cells were transduced with empty vector or RelB-IRES-GFP. Generated cells were FACS-sorted to obtain CD1a⁺GFP⁺ or CD1a^-GFP⁺ cell fractions, and cells were subsequently stimulated with CD40L for 24 hours. Cells were then analyzed for their capacity to stimulate allogeneic T cells. (A-C) Data are representative of at least 4 experiments.