Anti hmgb1

Male mice were anesthetized by intraperitoneal (i.p.) injection of 1% pentobarbital sodium solution (80 mg/kg, Biyuntian Institute of Biotechnology) and randomly assigned to the following groups: Control, LPS, positive control, and treatment group (n=4-6 per each group). Mice in the LPS and treatment groups then received an i.p., injection of LPS (15 mg/kg; Sigma-Aldrich, Merck KGaA) diluted in 200 µl sterile PBS. The control and positive control groups were administered only PBS by the same way and volume. The positive control and treatment groups received an intranasal (i.n.) injection of anti-HMGB1 (2.5 µg/g in 40 µl PBS, 2 h after LPS challenge; BioLegend, Inc.) (22 (link)) or rHMGB1 (0.5 µg/g in 40 µl PBS, 2 h after LPS challenge; R&D Systems, Inc.) (23 (link)-25 (link)), an i.p., injection of LPS from Rhodobacter sphaeroides (LPS-RS), a TLR2/4 antagonist (0.1 mg/mg in 200 µl endotoxin-free water, 1 h before LPS challenge; InvivoGen) (25 (link)-27 (link)) or FPS-ZM1, a RAGE antagonist (1.5 mg/kg in 200 µl PBS, 1 h before LPS challenge; EMD Millipore) (25 (link),28 (link)). All mice were sacrificed (cervical dislocation) 24 h after the last treatment and the blood, lung, bronchoalveolar lavage fluid (BALF), and spleen of mice were extracted for sample preparation.