LPS-Induced Lung Injury Mouse Model

LPS challenge was performed as previously described (de Souza Xavier Costa et al., 2017 (link)). Briefly, mice were exposed to nebulized LPS (Escherichia coli O111:B4, purified by phenol extraction; Sigma-Aldrich) at 0.8 mg/ml diluted in NaCl 0.9% (B Braun Medical, Sempach, Switzerland) in a plexiglas chamber connected to a nebulizer (System Assistance Medicale, Ledat, France) for 30 min. Control mice inhaled NaCl 0.9% only. Vehicle or ACT-1004-1239 was given p.o., 1 h prior (preventive setting) or 3 h post inhalation (therapeutic setting).
At different time points indicated in the figure legends (5, 24, 48, 72 h) following LPS or NaCl challenge, mice were euthanized with an overdose (150 mg/kg, intraperitoneally) of pentobarbital (Esconarkon, Streuli Pharma SA, Uznach, Switzerland), and samples were collected.