recombinant mouse il 17ra il 17r fc chimera protein  (R&D Systems)

Journal: Journal of Extracellular Vesicles

Article Title: An extracellular vesicular mutant KRAS‐associated protein complex promotes lung inflammation and tumor growth

doi: 10.1002/jev2.12307

Figure Lengend Snippet: EV‐KRAS complex facilitates inflammation through the IL‐17A/FGF21 axis. (a) HE staining of lung tissue from mice (blocked with IL‐17R/Fc or anti‐FGF21 antibody) intratracheally administered extracellular vesicle (EV) KRAS complex (liposome‐EV‐KRAS). (b) ELISA for TNFα, IL‐17A and IL‐6 from lung tissue from mice (blocked with IL‐17R/Fc or anti‐FGF21 antibody) intratracheal administered liposome‐EV‐KRAS. (c) Adenosine triphosphate (ATP) cell proliferation assay showing proliferation of Jurkat cells in response to lung tumour lysate with and without IL‐17A depletion using IL‐17A immunoprecipitation. (d) Luciferase assay for FGF21 promotor assessment, TC1 cells treated with respective lung tumour lysate with and without IL‐17A depletion using IL‐17A immunoprecipitation. (e) Western blot for PI3K, SIRT1, NFκB‐ TC1 cells treated with respective lung tumour lysate with and without FGF21 depletion using FGF21 immunoprecipitation. (f) qRT‐PCR for genes involved in EMT pathway‐ TC1 cells treated with respective lung tumour lysate with and without FGF21 depletion using FGF21 immunoprecipitation. For in vivo treatment mice were administered with 50 μl of 10 × 1010 liposome particles/ml and for in vitro treatment 2 × 108 liposome particle/ml for respective group were used. Data are mean ± SEM from three replicates using one‐way ANOVA, ** p < 0.01.

Article Snippet: In vivo cytokine neutralization experiments involved administering mAbs against FGF21 (Abclonal, Woburn, MA) and recombinant Mouse IL‐17RA/IL‐17R Fc Chimera Protein (R&D System) intraperitoneally.

Techniques: Staining, Enzyme-linked Immunosorbent Assay, Proliferation Assay, Immunoprecipitation, Luciferase, Western Blot, Quantitative RT-PCR, In Vivo, In Vitro

Journal: The Journal of Experimental Medicine

Article Title: Epithelial transglutaminase 2 is needed for T cell interleukin-17 production and subsequent pulmonary inflammation and fibrosis in bleomycin-treated mice

doi: 10.1084/jem.20101457

Figure Lengend Snippet: BLM-mediated pulmonary injury triggers a Th17 response. WT B6 and TG2 −/− mice received an intratracheal instillation of BLM (1.5 mg/kg) and were sacrificed to harvest BALF on the indicated days. Cells from BALF were stimulated with PMA and ionomycin and intracellular cytokines were detected by flow cytometric analysis. (A and B) Numbers of CD4 + cells producing IL-17 (A) or IFN-γ (B) in BALF of BLM-exposed WT B6 and TG2 −/− mice. (C) The percentages of CD4 + cells producing IL-17 or IFN-γ in BALF and draining lymph nodes (DLN) of WT B6 and TG2 −/− mice were determined by flow cytometric analysis 10 d after BLM exposure. Dot plots are gated on CD4 + T cells. (D–H) BLM-exposed WT B6 mice were treated with IL-17RA-Fc or isotype control for 10 d (100 µg/mouse/day, i.p.). IL-17 levels (D) and the numbers of CD4 + IL-17 + cells (E), total cells (F), and neutrophils (G) were determined by ELISA and flow cytometry, respectively. (H) BLM-exposed mice were treated with IL-17RA-Fc or isotype control for the times indicated. Quantitative analysis of the fibrotic area by H&E and Masson’s trichrome staining was performed 21 d after instillation of BLM. Data represent means ± SD of three independent determinations with BALF or lung tissues from n = 5 (H) or n = 3 mice/group for each time point (A–G).

Article Snippet: A proportion of the mice were injected intraperitoneally with CyM (40 mg/kg/day; Sigma-Aldrich), recombinant murine IL-17RA-Fc (100 µg/mouse/day; R&D Systems) or control human IgG1 (100 µg/mouse/day; R&D Systems) suspended in PBS.

Techniques: Enzyme-linked Immunosorbent Assay, Flow Cytometry, Staining