Bronchoalveolar lavage, lung tissue homogenates and histology.

Mice were euthanized by cervical dislocation under isoflurane inhalation and subjected to bronchoalveolar lavage (BAL) with PBS with 0.1% BSA. BAL fluid was clarified by centrifugation and stored at −80°C prior to analysis. Whole lung tissue isolated from mice was immersed in 1 mL PBS with 0.1% BSA and subjected to blade homogenization and clarification by centrifugation at 4°C. Clarified homogenates were stored at −80°C prior to analysis. For histology, lungs were perfused with 10 ml of PBS, inflated with 0.8 mL of ice cold 10% phosphate-buffered formalin and fixed in 10 mL cold 10% phosphate-buffered formalin. H&E-stained sections were prepared by Histoserv, Inc. (Germantown, MD, USA). Photographs of the microscopic images were obtained with a DMI4000 light microscope (Leica Microsystems, Wetzlar, Germany) equipped with a Retiga 2000R camera and analyzed using QCapture software (both from QImaging, Surrey, BC, Canada).