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. 2008 Mar 28;9(5):480–485. doi: 10.1038/embor.2008.37

Figure 5.

Figure 5

Suppression of synoviolin expression in synovial cells of mice with collagen-induced arthritis restores IRE1 protein expression. (A) SYVN1 siRNA expression plasmids or control (CTL) plasmids were transfected into synovial fibroblasts from normal and CIA mice. Three days after transfection, GFP-positive cells were sorted and the expression of SYVN1 (top panel) and IRE1 (middle panel) was determined by SYVN1 and IRE1 antibodies, respectively. The protein levels of actin were analysed as loading controls (bottom panel). (B,C) Synovial fibroblasts, transfected with either (B) control plasmids or (C) SYVN1 siRNA expression plasmids, were treated with or without tunicamycin (TM; 10 μg/ml) for 18 h. Apoptosis was determined by annexin V staining by using flow cytometry. (D) Synovial fibroblasts from normal and CIA mice were transfected with or without IRE1 expression plasmids. Three days after transfection, cells were treated with TM or dimethyl sulphoxide (DMSO) as a control. The percentages of apoptotic cells were analysed as described in (B) and (C). Error bars (mean±s.d.) represent data from three independent experiments. Student's t-test was used for statistical analysis. CIA, collagen-induced arthritis; IRE1, inositol-requiring enzyme 1; siRNA, short interfering RNA; SYVN1, synoviolin.