Mouse ears were treated with Aldara cream, containing 5% imiquimod, and 3 and 21 h later mice received into the ear pinna an injection of 12.5 μg LecB in 15 μl saline water. For DEC‐205 targeting, 1 μg anti‐DEC‐205‐A647 antibody diluted in 15 μl PBS was injected into ears pinna prior to Aldara cream application (Flacher et al, 2012 (link)). Auricular LNs were harvested 72 h afterwards. For single cell preparation, LNs were cut into small pieces and digested with 1 mg/ml collagenase D (Roche) and 0.1 mg/ml DNase I (Roche) in RPMI cell culture medium containing 2% FCS for 1 h at 37°C under agitation. For flow cytometry staining the antibodies were: CD11c‐PE‐Cy7, CD103‐PE (BD Pharmingen), MHC‐II‐AF700, CD205‐AF647 (BioLegend), and CD207 (Langerin)‐AF488 (Eurobio).