Immune Cell Profiling in Murine Lungs

Lungs were digested as previously described [10 (link)]. Cells were stained with antibodies: CD4-FITC, CD25-APC, FoxP3-PE from e-Bioscience (San Diego, CA, USA), CD4-PerCP-Cy5.5, CD45-APC-Cy7, Gr1-PerCP-Cy5.5, and CD11c-Brilliant Violet 605 from Biolegend (San Diego, CA, USA), CD3-PE-Cy7 and CD8-Alexa Fluor 700 from BD Bioscience (San Diego, CA, USA), CD68-FITC from AbD Serotec (Raleigh, NC, USA) and F4/80 from Life Technologies (Carlsbad, CA, USA). Dead cells were excluded using Live/Dead Fixable Blue Dead Cell Stain kit (Life Technologies, Carlsbad, CA, USA). Cells were analyzed using a LSR II flow cytometer (BD Biosciences, San Diego, CA, USA), and data were analyzed using Flow Jo 7.2 software (Tree Star, Ashland, OR, USA).