Anti il 5 trfk5

Murine lungs were chopped and incubated in RPMI1640 media with 1 mg/mL type IV collagenase (Worthington BioChem, Lakewood, NJ, USA) for at least 1 h 30 min at 37 °C. Red blood cells (RBC) were lysed using RBC lysis buffer (Biolegend, San Diego, CA, USA). Lung single cells were stained with following fluorochrome-conjugated antibodies. For surface staining, the following antibodies were used. Anti-CD45 (30-F11), Anti-CD3e (145-2C11), anti-CD11c (HL3), anti-CD11b (M1/70), anti-CD19 (ID3), anti-CD49b (DX5), anti-FcεRIα (MAR-1), anti-CD90,2 (30-H12), anti-F4/80 (BM8) and anti-Ly6G (1A8), purchased from Biolegend. Anti-SiglecF (E50-2440) was purchased from BD Bioscience (San Diego, CA, USA). For intracellular staining, the following antibodies were used: anti-IL5 (TRFK5) and anti-IL17A (TC11-18H10.1), purchased from Biolegend (San Diego, CA, USA). Anti-IL13 (eBio13A) was purchased from Thermo Fisher Scientific. For intracellular staining, a Fixation/Permeabilization Solution Kit with BD GolgiPlug (BD Biosciences, San Diego, CA, USA) was used following the manufacturer’s protocol. Flow cytometry was carried out using LSRFortessa™ X-20 (BD biosciences, San Jose, CA, USA) and analyzed by FlowJo (V10.2) software (BD biosciences, San Jose, CA, USA). The gating strategy for immune cell population was represented in Supplementary Figure S1.