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Mouse dendritic cells (DCs) were specifically activated in C57/Bl6 mice by subcutaneous injection of 50 μL of CD40 (1 mg/0.5 mL) or an isotype control (left images) into the left and right footpad. DC activation was analyzed in popliteal fossa lymph nodes after 72 hours. Cells were stained with CD11c-FITC, CD86-PE, CD205-Biotin/Anti-Biotin-APC, and CD3ε-PerCP antibodies, and analyzed by flow cytometry. The FcR Blocking Reagent was used to avoid Fc receptor–mediated antibody labeling. Cell debris, dead cells, and CD3+
T cells were excluded from the analysis based on scatter signals as well as propidium iodide and CD3ε-PerCP fluorescence.
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