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imm12173-sup-0001-figS1.tifimage/tif587KFigure S1. Phosphorylated IκB kinase (p-IKK) levels are decreased in suppressor of cytokine signalling 3 (SOCS3) knocked down M1 activated macrophages. Macrophages were transfected with control short interfering RNA (siRNA) or SOCS3 siRNA. 48 hr following transfection, cells were stimulated without or with lipopolysaccharide (LPS) for 0, 15 and 30 min and p-IKK and IKK levels were assessed by Western blotting. Densitometric analysis was performed on scanned bands and results presented as the ratio of p-IKK to β-actin in arbitrary units (a.u.). Figure is representative of five individual experiments.
imm12173-sup-0002-FigS2.tifimage/tif928KFigure S2. Human monocyte derived macrophages were activated with 100 ng/ml lipopolysaccharide (LPS) for 4 hr or left untreated (non-stimulated). Total macrophage protein was isolated and expression of protein and loading control β-actin detected by Western blot and normalized by densitometric analysis of scanned bands in arbitrary units (= 4). (a) Suppressor of cytokine signalling 3 (SOCS3) expression increases upon LPS stimulation in human monocyte derived macrophages. (b) SOCS3 knockdown (S3) in human monocyte derived macrophages decreases SOCS protein expression as compared with control (con) short interfering RNA (siRNA) transfected cells. (c) Knockdown of SOCS3 (S3) in human monocyte derived macrophages increases phosphorylated signal transducer and activator of transcription 3 (pSTAT3), but not pSTAT1, levels and (d) increases SOCS1 expression after LPS stimulation.

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