On day 21, mice were administered DiO-labeled liposomes via intratracheal injection and sacrificed after 24 h. The lungs of the mice were minced into a homogenous paste and treated with 3 ml of 1 mg/ml collagenase 1 diluted in HBSS. After incubation in a 15 ml centrifuge tube for 1.5 h at 37 °C in a table concentrator, the cell suspensions were passed through a 100 µm Falcon nylon cell strainer (Corning, NY, USA) and resuspended in stain buffer (PBS containing 2% FBS). Antibodies against F4/80-VioBlue, CD11c-PE and CD11b-APC were obtained from BD Biosciences Pharmingen. Staining of 100 µl cell suspensions containing 1×106 cells was performed in stain buffer at 4 °C for 30 min. The samples were washed twice prior to flow cytometry analysis.

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