Il 1ra

The IL-1RA (Cayman Chemical, Ann Arbor, MI) was administered daily at 0.2 μg per mouse (approximately 0.08 mg/kg) via intranasal instillation (volume 5 μL) starting on the day of RSV infection for seven doses. This dose was chosen based on prior dose-finding experiments that suggested this dose was safe and potentially effective.

Mouse monoclonal anti-actin (AM1021b) was purchased from Abgent Biotechnology (San Diego, CA). Rabbit polyclonal anti-TRPA1 (Western blotting: PA1-46159, Invitrogen; immunofluorescence: ACC-037, Alomone). Rabbit polyclonal anti-phospho-ERKI/2 (AP3906A, Abgent), Mouse polyclonal anti-total ERK1/2 (AM2189B, Abgent), and mouse monoclonal anti-fibronectin (F6140, MilliporeSigma) were used. HRP-conjugated antirabbit and antimouse secondary antibodies were purchased from Thermo Scientific (Waltham, MA) and Abcam, respectively. Cross-adsorbed Alexa Fluor 555 goat antirabbit immunoglobulin was purchased from Life Technologies (Grand Island, NY). Highly cross-adsorbed CF555 donkey antimouse IgG (catalog # 20037) was purchased from Biotium, Inc (Fremont, CA). Unless otherwise specified, all reagents were purchased from MilliporeSigma (Burlington, MA, USA). IL-1β (ProSpec-Tany TechnoGene Ltd, East Brunswick, NJ USA) and IL-1α (Sino Biological, Life Technologies, MD, USA) were used. IL-1RA, OAG, U-0126, A-967079, and thapsigargin (Cayman Chemical; Ann Arbor, MI USA), polygodial, and A-784168 (Tocris Bioscience, Bristol, UK), BAPTA (Assay BioTech, Sunnyvale, CA USA) were used. Lyophilized IL-1β was reconstituted in PBS +0.1% BSA. Control cells were treated with PBS +0.1% BSA.