Skip to main content
. Author manuscript; available in PMC: 2018 Oct 1.
Published in final edited form as: Nat Immunol. 2017 Aug 28;18(10):1084–1093. doi: 10.1038/ni.3821

Figure 1.

Figure 1

Mouse and human neutrophils are responsive to IFN-λ. (a) qPCR analysis of Ifnlr1 mRNA (left) and Il10rb mRNA (right), which encode the two components of the IFN-λ receptor, in mouse bone-marrow-derived neutrophils (PMN), primary intestinal epithelial cells (IEC), splenic plasmacytoid DCs (pDC) and splenic conventional DCs (cDC) derived from wild-type (WT) or IFNLR1-deficient (Ifnlr1−/−) mice (key); results are presented relative to those of the control gene Gapdh. ND, not detectable. (b) qPCR analysis of Viperin mRNA in neutrophils (left), plasmacytoid DCs (middle) or conventional DCs (right) left unstimulated (US) or stimulated for 3 h with IFN-λ or IFN-β (100 U/ml each) (key); results presented as in a. (c) qPCR analysis of Ifnlr1 in cells as in a, as well as T cells, B cells and peritoneal macrophages (pM0), left unstimulated or stimulated for 3 h with LPS (10 μg/ml) or TNF (100 ng/ml) (key). (d) qPCR analysis of Ifnlr1 (left) in human neutrophils left unstimulated or stimulated for 3 h with LPS (10 μg/ml), and of Viperin mRNA (right) in human neutrophils treated for 3 h with IFN-λ2 (100 U/ml) or IFN-β (100 U/ml) (horizontal axis) and left unstimulated or stimulated for 3 h with LPS (10 μg/ml) (key). (e) Immunoblot analysis of phosphorylated (p−) STAT1 and STAT3, and β-actin (loading control throughout), in neutrophils derived from the bone marrow of wild-type mice (top) or Ifnlr1−/− mice (bottom) and stimulated for 0–60 min (above lanes) with IFN-λ or IFN-β (blots cropped to show bands of the appropriate molecular size). (f) NanoString analysis of gene expression (nCounter Mouse Inflammation v2 panel; genes modulated more than twofold, with a P value of > 0.05) in mouse neutrophils stimulated for 30 min, 1 h or 3 h (left margin) with IFN-λ or IFN-β (100 U/ml). Data are from one of three experiments (a; mean + s.e.m. of three biological replicates) or one experiment representative of three experiments (b,d; mean + s.e.m. of three biological replicates) or are representative of three independent experiments (c,e,f; mean + s.e.m. in c).