Legendplex mouse anti virus response standard panel

Bronchoalveolar lavage was carried out by cannulating mouse tracheas with a catheter and flushing the air space with saline three times. The levels of IFN-γ, TNF-α, IL-6, IL-10, IL-12p70, and MCP-1 in the BALf was determined using a BD Cytometric Bead Array Mouse Inflammation Kit (BD Biosciences, San Diego, CA, USA) according to the manufacturer's instructions. Samples were then analyzed on a Becton Dickinson FACSCanto flow cytometer (Franklin Lakes, NJ, USA) and data analyzed using FlowJo software (Tree Star, Inc., Ashland, OR, USA). In some experiments a cytokine/chemokine concentrations in bronchial alveolar lavage fluid (BALf) was measured using a LEGENDplex mouse anti-virus response standard panel (Biolegend) following manufacturer's instructions.

For collection of the BALF, the trachea was exposed and cannulated, and the lungs were lavaged three times with 300 μl of PBS. Cytokine/chemokine concentrations in BALF were measured using LEGENDplex mouse antivirus response standard panel (BioLegend) following the manufacturer’s instructions. Samples were acquired using a Becton Dickinson FACSCanto II flow cytometer, and data were analyzed using the FlowJo software package (Tree Star Inc., Ashland, OR, USA).