recombinant human srage  (R&D Systems)

Journal: Frontiers in Immunology

Article Title: DNA-Mediated Interferon Signature Induction by SLE Serum Occurs in Monocytes Through Two Pathways: A Mechanism to Inhibit Both Pathways

doi: 10.3389/fimmu.2018.02824

Figure Lengend Snippet: ISG induction by SLE serum requires either RAGE or FcRlla. (A) Primary human monocytes were incubated with serum from different SLE patients, B2, M91, B31, F72, or control sera, for 4 h. ISG expression was evaluated using qPCR. Data plotted represents fold induction for each gene relative to the sample incubated with control serum and is representative of 30 samples tested. (B) RAGE and FcRlla levels in monocytes treated with RAGE or FcRIIa siRNA, assessed by qPCR. (C) Monocytes were transfected with RAGE, FcRlla, or control siRNA, prior to incubation with SLE serum for 4 h. For each sample (control, RAGE, or FcRlla siRNA transfected cells) fold induction of ISGs was calculated as the ratio of gene expression between treated and untreated cells. (D) Primary human monocytes were treated with SLE serum, with or without sRAGE or monoclonal anti-FcRlla antibody for 4 h. ISG expression was analyzed by qPCR. Results in each case (B–D) indicate mean ± SD of three independent experiments. * p < 0.05; ** p < 0.01; *** p < 0.001.

Article Snippet: Human recombinant sRAGE was purchased from ProSpec (East Brunswick NJ, USA).

Techniques: Incubation, Control, Expressing, Transfection, Gene Expression

Journal: Frontiers in Immunology

Article Title: DNA-Mediated Interferon Signature Induction by SLE Serum Occurs in Monocytes Through Two Pathways: A Mechanism to Inhibit Both Pathways

doi: 10.3389/fimmu.2018.02824

Figure Lengend Snippet: (A) Biacore T200 analysis of DWEYS inhibition of HMGB1-RAGE binding. sRAGE was immobilized on a CM5 sensor chip, HMGB1 with or without varying concentrations of DWEYS were added as analytes. IC50 was 34.7 μM. (B) Inhibition curve used for IC50 evaluation as noted in the methods section. (C,D) Human monocytes were incubated with labeled HMGB1 (red) alone ( C , top panel) or HMGB1 and DWEYS ( C , middle panel) or HMGB1 and control peptide ( C , bottom panel) for 20 min. Cells were counterstained with DAPI and imaged using a confocal laser scanning microscope. (D) Indicates adjusted fluorescence intensity for three experiments. (E) Primary human monocytes were incubated with HMGB1 and with or without DWEYS or control peptide for 4 h. lSG expression was evaluated using qPCR. Results indicate mean ± SD of three independent experiments. * p < 0.05.

Article Snippet: Human recombinant sRAGE was purchased from ProSpec (East Brunswick NJ, USA).

Techniques: Inhibition, Binding Assay, Incubation, Labeling, Control, Laser-Scanning Microscopy, Fluorescence, Expressing

Journal: Scientific Reports

Article Title: Inhibition of the Receptor for Advanced Glycation End-Products in Acute Respiratory Distress Syndrome: A Randomised Laboratory Trial in Piglets

doi: 10.1038/s41598-019-45798-5

Figure Lengend Snippet: RAGE inhibition restores alveolar fluid clearance after acid-induced lung injury. Measurement of net alveolar fluid clearance (AFC) rate as a marker of epithelial function in uninjured (Sham), acid-injured (HCl) and acid-injured piglets treated with RAGE antagonist peptide (HCl + RAP) or recombinant sRAGE (HCl + sRAGE) (n = 12 per group at each time point). Values are reported as box and whisker plots. The Kruskal-Wallis test, with post-hoc Bonferroni test for pairwise comparisons were used. P = 0.02 for the overall multiple group test; no statistical difference was found in post-hoc tests.

Article Snippet: The “sRAGE group” (n = 12) included animals with HCl-induced lung injury that also received intravenous treatment with sRAGE (3 mg.kg −1 ) (Recombinant Human RAGE Fc Chimera, R&D Systems, Minneapolis, MN).

Techniques: Inhibition, Marker, Recombinant, Whisker Assay

Journal: Scientific Reports

Article Title: Inhibition of the Receptor for Advanced Glycation End-Products in Acute Respiratory Distress Syndrome: A Randomised Laboratory Trial in Piglets

doi: 10.1038/s41598-019-45798-5

Figure Lengend Snippet: RAGE inhibition improves arterial oxygenation after acid-induced lung injury. Arterial oxygen tension (PaO 2 )/inspiratory oxygen fraction (FiO 2 ) in uninjured (Sham), acid-injured (HCl) and acid-injured piglets treated with RAGE antagonist peptide (HCl + RAP) or recombinant sRAGE (HCl + sRAGE) (n = 12 per group at each time point). Individual values are reported. Time × group interaction and post-hoc comparisons were verified using random effects models to analyse longitudinal evolution of variables.

Article Snippet: The “sRAGE group” (n = 12) included animals with HCl-induced lung injury that also received intravenous treatment with sRAGE (3 mg.kg −1 ) (Recombinant Human RAGE Fc Chimera, R&D Systems, Minneapolis, MN).

Techniques: Inhibition, Recombinant

Journal: Scientific Reports

Article Title: Inhibition of the Receptor for Advanced Glycation End-Products in Acute Respiratory Distress Syndrome: A Randomised Laboratory Trial in Piglets

doi: 10.1038/s41598-019-45798-5

Figure Lengend Snippet: RAGE inhibition decreases alveolar-capillary permeability after acid-induced lung injury. Values are reported as box and whisker plots. ( A ) Level of total protein in the bronchoalveolar lavage (BAL) fluid from uninjured (Sham), acid-injured (HCl) and acid-injured piglets treated with RAGE antagonist peptide (HCl + RAP) or recombinant sRAGE (HCl + sRAGE) (n = 12 per group). The Kruskal-Wallis test, with post-hoc Bonferroni test for pairwise comparisons were used. ( B ) Extravascular lung water, as measured by transpulmonary thermodilution (Picco + , Pulsion SA) and indexed to body weight, in uninjured (Sham), acid-injured (HCl) and acid-injured piglets treated with RAGE antagonist peptide (HCl + RAP) or recombinant sRAGE (HCl + sRAGE) (n = 12 per group at each time point). Time × group interaction and post-hoc comparisons were verified using random effects models to analyse longitudinal evolution of variables.

Article Snippet: The “sRAGE group” (n = 12) included animals with HCl-induced lung injury that also received intravenous treatment with sRAGE (3 mg.kg −1 ) (Recombinant Human RAGE Fc Chimera, R&D Systems, Minneapolis, MN).

Techniques: Inhibition, Permeability, Whisker Assay, Recombinant

Journal: Scientific Reports

Article Title: Inhibition of the Receptor for Advanced Glycation End-Products in Acute Respiratory Distress Syndrome: A Randomised Laboratory Trial in Piglets

doi: 10.1038/s41598-019-45798-5

Figure Lengend Snippet: RAGE inhibition decreases alveolar inflammation after acid-induced lung injury. Measurement of bronchoalveolar lavage (BAL) levels of ( A ) tumour necrosis factor (TNF)-α, ( B ) interleukin (IL)-6, ( C ) IL-1β and ( D ) IL-18 in uninjured (Sham), acid-injured (HCl) and acid-injured piglets treated with RAGE antagonist peptide (HCl + RAP) or recombinant sRAGE (HCl + sRAGE) (n = 12 per group). Values are reported as box and whisker plots. The Kruskal-Wallis test, with post-hoc Bonferroni test for pairwise comparisons were used.

Article Snippet: The “sRAGE group” (n = 12) included animals with HCl-induced lung injury that also received intravenous treatment with sRAGE (3 mg.kg −1 ) (Recombinant Human RAGE Fc Chimera, R&D Systems, Minneapolis, MN).

Techniques: Inhibition, Recombinant, Whisker Assay

Journal: Scientific Reports

Article Title: Inhibition of the Receptor for Advanced Glycation End-Products in Acute Respiratory Distress Syndrome: A Randomised Laboratory Trial in Piglets

doi: 10.1038/s41598-019-45798-5

Figure Lengend Snippet: (A ) RAGE inhibition decreases histological features of lung injury. Lung injury scores were higher in acid-injured (HCl) than in uninjured piglets (Sham) and acid-injured piglets treated with RAGE antagonist peptide (HCl + RAP) or recombinant sRAGE (HCl + sRAGE) (n = 12 per group). Values are reported as box and whisker plots. The Kruskal-Wallis test, with post-hoc Bonferroni test for pairwise comparisons were used. (B – E) Representative hematoxylin and eosin–stained sections at x20 original magnification of (B) uninjured piglets (Sham), (C) acid-injured piglets (HCl), acid-injured piglets treated with (D) RAGE antagonist peptide (HCl + RAP) or (E) recombinant sRAGE (HCl + sRAGE). There was greater cellularity consisting mainly of neutrophils (black arrowheads), more alveolar wall thickening (white arrowheads), and more areas of atelectasis and increased alveolar disruption, proteinous debris, and hemorrhage (black arrows) in untreated, acid-injured than in uninjured or treated piglets. Scale bars, 100 µm.

Article Snippet: The “sRAGE group” (n = 12) included animals with HCl-induced lung injury that also received intravenous treatment with sRAGE (3 mg.kg −1 ) (Recombinant Human RAGE Fc Chimera, R&D Systems, Minneapolis, MN).

Techniques: Inhibition, Recombinant, Whisker Assay, Staining, Disruption