Ifnλ2

Neutrophils from the lung, spleen, and bone marrow as well as bone marrow-derived T and B cells were isolated from naïve wild-type, IFNAR^−/−, IFNLR1^−/−, IFNAR^−/−IFNLR1^−/−(DKO), MRP8^cre Ifnlr1^fl/fl, MRP8^cre Stat1^fl/fl, MRP8^cre, Ifnlr1^fl/fl, and Stat1^fl/fl mice. Neutrophils and B cells were stimulated with 100 ng of IFNα2 (Novoprotein) or IFNλ2 (PBL). B cells were sorted as Live CD45⁺B220⁺, T cells were sorted as Live CD45⁺Thy1.2⁺, and neutrophils were sorted as Live CD45⁺CD11b⁺Ly6C^intLy6G⁺. These populations were sorted to >99% purity. Cell lysates were collected in lysis buffer containing protease and phosphatase inhibitors. Equal amounts of total protein was separated on 7.5% SDS-PAGE gels, transferred to Nitrocellulose 0.45 µm membrane (BIO-RAD), and subsequently probed with antibody against phosphorylated STAT1 (BD #612133) and β-actin (Sigma #A5441). For PCR genotyping, DNA from sorted cells was isolated using Qiagen DNeasy kit, and run on a 4% agarose gel. Control DNA was extracted from tail snips.