DNGR-1⁺ BDCA3⁺ DCs are found in nonlymphoid tissues of humanized mice. (A) hCD11c⁺ HLA-DR⁺ lung (top left panel) or kidney (bottom left panel) cells were analyzed as shown for the expression of BDCA3 versus CD11b (right panel), after having gated out Dump⁺ auto⁺ cells (middle panels). Two populations were defined: gate I, BDCA3⁺, CD11b⁻; and gate II, BDCA3⁻, CD11b⁺. The Dump channel contains the following stainings grouped together: a Lin cocktail (anti–human CD3/CD19/CD56), an anti–mouse CD45, and a viability marker. (B) Lung (top panels) and kidney (bottom panel) BDCA3⁺ (left panels) or CD11b⁺ (right panels) cells, as defined in panel A by gates I and II, were analyzed for the expression of DNGR-1 (black) or for an isotype control Ab (grey). (C) hCD11c⁺ HLA-DR⁺ BDCA3⁺ lung cells, as gated in panel A, expressing DNGR-1⁺ (top left) or not (top right) were analyzed for CD103 expression (black) or for an isotype control antibody (grey). As an internal control, total lung cells were also analyzed for these markers versus the Dump channel (bottom). Numbers indicate percentage of cells in each of the indicated gates. Arrows show gating strategy. (A-B) Data are representative of at least 3 independent experiments.