DCs from CTX-treated mice are more potent at antigen presentation. Whole CD11c⁺ DCs from CTX-treated or naive mice were sorted from LNs with anti-CD11c magnetic-activated cell sorting beads. (A) Graded doses of DCs were cocultured with allogeneic T cells from Balb/c mice or were pulsed with OVA peptide and cocultured with antigen-specific CD8⁺ T cells from OT-I mice. During the incubation, MTS/PMS solution was added to the culture to measure the proliferation of T cells. Absorbance at 490 nm was recorded using an enzyme-linked immunosorbent assay plate reader. Representative data from 3 independent experiments are shown. (B) DCs from CTX-treated or naive mice were sorted as described for panel A and cultured in vitro with granulocyte-macrophage colony-stimulating factor for 20 hours with or without stimulation (LPS + IFN-γ). Brefeldin A was present during the last 12 hours of the incubation. (C) DCs were stained with anti-CD11c, MHC class II, B220, Gr-1, and CD8 to identify different DC subsets as defined in Figure 1A and examined for production of IL-12 p40/70. Baseline levels and augmentation of IL-12 p40/70 with the addition of LPS and IFN-γ are depicted. Data are representative of 3 independent experiments. *Statistically significant.