DCs carry model antigen to airway-adjacent regions as well as LNs. (A) Scheme for allergen challenge includes three i.p. injections of OVA/alum and three i.n. instillations of OVA or PBS (black arrows), an i.n. instillation of red beads (red arrow), and an i.n. instillation of blue beads (blue arrow) to dissect uptake versus trafficking. (B) Example images of beads in CD11c-EYFP DCs. Arrows indicate example beads in DCs. Bars, 200 µm. (C) Quantification of beads in CD11c-EYFP DCs yielded the ratio of equilibrated (beads inhaled 48 h before analysis) to immediate (beads inhaled directly before analysis). The airway region is defined as 75 µm from the inner airway surface. (D) CD103, CD11b, and LN-resident (LNR) DC populations were compared by FACS after PBS or OVA challenge. (E) The phenotype of DCs carrying model antigen to the LN was analyzed by FACS 48 h after bead inhalation. (C–E) Error bars represent SEM. All data represents at least eight mice per group from at least five independent experiments.