Fig. 2

Cytokines preferentially enhance VCAM1, VEGF, and ROS expression in RA FLS compared to OA FLS. RA and OA FLS were stimulated with or without IL-17 (10 ng/ml) and TNF-α (10 ng/ml) for 1 h. Following replacement of culture media, flow cytometry analysis was performed after 23 h with the indicated antibodies. a Data represent the mean fluorescence intensity (M.F.I.) for each factor. b Levels of secreted VEGF in stimulated and unstimulated RA and OA FLS were measured using an ELISA. c Mitochondria-specific ROS (mROS) were detected by MitoSox dye at the indicated time points after cytokine stimulation. Results are presented as mean. d The unstimulated control is represented by the dotted line, and cytokine-stimulated cells are represented by the bold line. e ROS levels in RA and OA FLS at 24 h are shown. Data are shown as fold change of the M.F.I. in stimulated cells compared to unstimulated control cells. Each symbol represents an individual donor. Each group contains five donors. f Following treatment with 20 μM MitoTEMPO for 1 h, RA FLS were stimulated with IL-17 (10 ng/ml) and TNF-α (10 ng/ml) for 1 h. Cell migration was measured using a transwell chamber after 23 h and staining with crystal violet dye. g Following solubilization, the crystal violet dye was quantitated. The bar represents the mean. Data represent one experiment, which was performed in triplicate with similar results. *p < 0.05. Scale bar, 100 μm. Magnification is × 100