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Figure 2 | Arthritis Research & Therapy

Figure 2

From: Cyr61 is involved in neutrophil infiltration in joints by inducing IL-8 production by fibroblast-like synoviocytes in rheumatoid arthritis

Figure 2

Cyr61 promotes IL-8 production in RA FLS. (A) IL-8 and Cyr61 levels in SF from RA patients (n = 21) and OA patients (n = 11) were detected by ELISA. (B) IL-8 mRNA expression in RA FLS stimulated by Cyr61 (0.62, 1.25, 2.5, 5, 10 μg/ml) for two hours was determined by real-time PCR using a housekeeping gene as an endogenous control. (C) IL-8 mRNA expression in RA FLS stimulated by 5 μg/ml Cyr61 (0, 2, 4, 6, 8 hours) was determined by real-time PCR. (D) The concentration of IL-8 in supernatant was evaluated by ELISA. FLS were seeded into 24-well plates and stimulated with exogenous Cyr61 (0.62, 1.25, 2.5, 5, 10 μg/ml) for 48 hours. (E) The expression of IL-8 mRNA and protein in Cyr61-knockdown FLS was detected by real-time PCR and ELISA, respectively. FLS were transfected with siCyr61 or siNC as described in Methods. After siRNA transfection, FLS were stimulated with 5 μg/ml Cyr61 for 2 hours and analyzed for IL-8 expression by real-time PCR and supernatants from FLS stimulated with 5 μg/ml Cyr61 for 48 hours were analyzed for IL-8 production by ELISA. (F) Cyr61-stimulated production of IL-8 by FLS was inhibited by 093G9. FLS were pretreated with 20 μg/ml anti-Cyr61 monoclonal antibody (named 093G9) or control IgG for one hour. After pretreatment, FLS were stimulated with 5 μg/ml Cyr61 for 48 hours and supernatants were analyzed for IL-8 production by ELISA. Data represent the mean ± SEM of at least three independent experiments. * P <0.05, ** P <0.01. FLS, fibroblast-like synoviocytes; OA, osteoarthritis; RA, rheumatoid arthritis; SEM, standard error of the mean; siNC, control siRNA.

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