Twenty-four hours after LPS challenge, animals were weighted, anaesthetized with isoflurane and sacrificed by bleeding from the abdominal aorta for bronchoalveolar lavage fluid (BALF) collection, performed as previously described (Nassini et al. 2012 (link)). Bronchoalveolar lavage (BAL) fluid supernatants were frozen at −80°C for simultaneous quantitation of multiple cytokines/chemokines using a Bio-Plex™ Cytokine Assay Kit (Bio-Rad Laboratories, Segrate, Milano, Italy).
The cell pellet was resuspended in 0.2 mL of PBS and cell counts were obtained using a particle counter (Dasit XT 1800J, Cornaredo, Milano, Italy).
The cell pellet was resuspended in 0.2 mL of PBS and cell counts were obtained using a particle counter (Dasit XT 1800J, Cornaredo, Milano, Italy).
