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Surface expression of receptors for IL-31 and IL-33 on human eosinophils and fibroblasts.

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posted on 20.02.2013, 09:48 by Chun-Kwok Wong, Karen Ming-Lam Leung, Huai-Na Qiu, Joyce Yin-Sau Chow, Angela On Kei Choi, Christopher Wai-Kei Lam

(A) Surface expression of OSMR-β, IL-31RA and ST2 on eosinophils, peripheral blood mononuclear cells (PBMC), neutrophils and fibroblasts (5×105 cells) was determined Western blot. Triplicate experiments were performed with essentially identical results and representative figure is shown. Neutrophils and PBMC were served as cell controls, and β-actin was used as protein control to ensure an equal amount of loaded protein. (B) Surface expression of ST2 on fibroblasts was determined by flow cytometry. Results are expressed as representative histogram of relative cell counts with mean fluorescence intensity (MFI). (C) Functional activity of the receptor complex for IL-31 and IL-33. IL-31 or IL-33 (50 ng/ml) was added to eosinophils or fibrolasts (5×105 cells) for 10 min. Phosphorylation of STAT3 (pSTAT3) in eosinophils and fibroblasts was determined by flow cytometry. Results are shown in MFI subtracting corresponding isotypic control and expressed as the arithmetic mean plus SD of three independent experiments in bar chart. Representative histograms illustrate the intracellular expression of pSTAT3 upon IL-31 or IL-33 stimulation in permeabilized eosinophils and fibroblasts. *p<0.05 comparing with corresponding medium control. EOS: eosinophils; NEU: neutrophils; FIB: dermal fibroblasts.