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. Author manuscript; available in PMC: 2010 Nov 4.

Published in final edited form as: J Immunol. 2006 Jan 15;176(2):711–715. doi: 10.4049/jimmunol.176.2.711

IL-17RA constructs used in this study. A, Schematic diagram of IL-17RA and TNFR FRET constructs. Full-length and truncated murine IL-17RA cD-NAs were fused in-frame to YFP or CFP. A truncated TNFR p60 fused to YFP was used as a control. IL-17R constructs have N-terminal FLAG tags, and the TNFR construct has an N-terminal hemagglutinin tag. TM, transmembrane domain, STIR, similar expression of fibroblast growth factor receptor/Toll/IL-1 receptor domain. B, IL-17R/CFP and IL-17R/YFP deliver normal IL-17-dependent signals. IL-17R-deficient murine fibroblasts were transiently transfected with a reporter construct containing the murine 24p3 proximal promoter upstream of luciferase (25) and an internal Renilla-luciferase plasmid along with a vector control (□), wild-type IL-17RA (■), or IL-17R/CFP and IL-17R/YFP constructs (▨ and , respectively). Cells were stimulated with IL-17 (100 ng/ml) and/or TNF-α (2 ng/ml) for 6 h and luciferase assays were performed in triplicate. SDs are shown. C, Normal cell surface expression of IL-17R (1-441). HEK293.IL-17R(1-441)/CFP and/YFP cells (black line), ST2 cells (gray line), or IL-17RA-deficient fibroblasts (solid histogram) were stained with anti-mouse IL-17RA-PE.

Inline graphic — IL-17RA constructs used in this study. A, Schematic diagram of IL-17RA and TNFR FRET constructs. Full-length and truncated murine IL-17RA cD-NAs were fused in-frame to YFP or CFP. A truncated TNFR p60 fused to YFP was used as a control. IL-17R constructs have N-terminal FLAG tags, and the TNFR construct has an N-terminal hemagglutinin tag. TM, transmembrane domain, STIR, similar expression of fibroblast growth factor receptor/Toll/IL-1 receptor domain. B, IL-17R/CFP and IL-17R/YFP deliver normal IL-17-dependent signals. IL-17R-deficient murine fibroblasts were transiently transfected with a reporter construct containing the murine 24p3 proximal promoter upstream of luciferase (25) and an internal Renilla-luciferase plasmid along with a vector control (□), wild-type IL-17RA (■), or IL-17R/CFP and IL-17R/YFP constructs (▨ and , respectively). Cells were stimulated with IL-17 (100 ng/ml) and/or TNF-α (2 ng/ml) for 6 h and luciferase assays were performed in triplicate. SDs are shown. C, Normal cell surface expression of IL-17R (1-441). HEK293.IL-17R(1-441)/CFP and/YFP cells (black line), ST2 cells (gray line), or IL-17RA-deficient fibroblasts (solid histogram) were stained with anti-mouse IL-17RA-PE.