Journal: Scientific Reports

Article Title: SCARF-1 promotes adhesion of CD4 + T cells to human hepatic sinusoidal endothelium under conditions of shear stress

doi: 10.1038/s41598-017-17928-4

Figure Lengend Snippet: In vitro expression of SCARF-1 in HSEC can be up-regulated by proinflammatory cytokines and LPS. ( a ) Representative images of immunofluorescent staining of SCARF-1 (green) with DAPI nuclear stain (blue). Scale bar = 25 µm. ( b ) Representative Western blot ( left panel ) and quantification ( right panels ) of the 180 kDa (dimeric) and 90 kDa (monomeric) species of SCARF-1 in stimulated HSEC compared to media alone control (Con). Results are representative of 3 independent experiments and are regions cropped from the same membrane (see Supplementary Figure ). ( c ) Fold change in SCARF-1 protein expression measured by cell-based ELISA in stimulated HSEC. ( d ) qPCR analysis of SCARF-1 mRNA in stimulated HSEC. ( a – d ) HSEC were treated with 10 ng/ml of tumour necrosis factor (TNF)α, 10 ng/ml of interferon (IFN)γ, or both in combination or with 1 µg/ml of LPS for 24 h. ( c and d ) Dotted lines indicate control level of expression. * and ** indicate statistical significance where p ≤ 0.05 and p ≤ 0.01, respectively. ( b and d ) n = 3 and ( c ) n = 5 independent experiments with different HSEC donors in each group.

Article Snippet: Prior to stimulation, HSEC were incubated with endothelial SFM with 10% FCS for 24 h. HSEC were then stimulated with either proinflammatory cytokines (tumour necrosis factor (TNF)α and recombinant human interferon (IFN)γ; 10 ng/ml; both from PeproTech), lipopolysaccharide (LPS) from E . coli 0111:B4 (1 μg/ml; Sigma-Aldrich) or tumourigenic growth factors (VEGF and HGF; 10 ng/ml) for 24 h.

Techniques: In Vitro, Expressing, Staining, Western Blot, Control, Membrane, In-Cell ELISA

Journal:

Article Title: Role of JAK2 signal transductional pathway in activation and survival of human peripheral eosinophils by interferon-gamma (IFN-?)

doi: 10.1046/j.1365-2249.1999.01068.x

Figure Lengend Snippet: Effect of IL-5, IFN-γ and tumour necrosis factor-alpha (TNF-α) on eosinophil survival. Purified eosinophils were cultured in the presence of rhIL-5 (10 ng/ml; •), rhIFN-γ (1000 U/ml; □), rhTNF-α (10 ng/ml; ○) or medium only (▪) for 48 h. The viability of eosinophils was determined by examining 200 cells stained with trypan blue dye exclusion.

Article Snippet: Eosinophils (5 × 10 5 ) with recombinant human IFN-γ (rhIFN-γ; Genzyme, Boston, MA), rhIL-5 (Genzyne), or recombinant human tumour necrosis factor-alpha (rhTNF-α; PharMingen, San Diego, CA) were resuspended in the 1 ml of culture medium at 37°C up to 48 h in a humidified 5% CO 2 in air.

Techniques: Purification, Cell Culture, Staining

Journal:

Article Title: Role of JAK2 signal transductional pathway in activation and survival of human peripheral eosinophils by interferon-gamma (IFN-?)

doi: 10.1046/j.1365-2249.1999.01068.x

Figure Lengend Snippet: Effect of JAK2 inhibitor on IFN-γ-induced eosinophil survival. A JAK2 inhibitor, AG-490 (10−5–5 × 10−5 m) was added to the eosinophil culture medium with rhIL-5 (10 ng/ml), rhIFN-γ (1000 U/ml), or recombinant human tumour necrosis factor-alpha (rhTNF-α; 10 ng/ml). The viability of 48 h cultured eosinophils was determined by examining 200 cells stained with trypan-blue dye exclusion.

Article Snippet: Eosinophils (5 × 10 5 ) with recombinant human IFN-γ (rhIFN-γ; Genzyme, Boston, MA), rhIL-5 (Genzyne), or recombinant human tumour necrosis factor-alpha (rhTNF-α; PharMingen, San Diego, CA) were resuspended in the 1 ml of culture medium at 37°C up to 48 h in a humidified 5% CO 2 in air.

Techniques: Recombinant, Cell Culture, Staining

Journal:

Article Title: Role of JAK2 signal transductional pathway in activation and survival of human peripheral eosinophils by interferon-gamma (IFN-?)

doi: 10.1046/j.1365-2249.1999.01068.x

Figure Lengend Snippet: Effect of JAK2 inhibitor on IFN-γ-induced CD69 expression on 24 h cultured eosinophils. A JAK2 inhibitor, AG-490 (10−5–5 × 10−5 m) was added to the eosinophil culture medium with rhIL-5 (10 ng/ml), rhIFN-γ (1000 U/ml), or recombinant human tumour necrosis factor-alpha (rhTNF-α; 10 ng/ml) from time zero to 24 h. Flow cytometric analysis was then performed. The data on mean fluorescence intensity (MFI) of CD69 are shown in ratio form as described in Materials and Methods. Data are mean ± s.d. for six experiments. Significantly different from the mean value of the control: *P < 0.05; **P < 0.01.

Article Snippet: Eosinophils (5 × 10 5 ) with recombinant human IFN-γ (rhIFN-γ; Genzyme, Boston, MA), rhIL-5 (Genzyne), or recombinant human tumour necrosis factor-alpha (rhTNF-α; PharMingen, San Diego, CA) were resuspended in the 1 ml of culture medium at 37°C up to 48 h in a humidified 5% CO 2 in air.

Techniques: Expressing, Cell Culture, Recombinant, Fluorescence