Fig. 5.

Regulation of expression of proinflammatory cytokines, migration, and invasion of rheumatoid arthritis (RA) fibroblast-like synoviocytes (FLSs) by mucin 1 (MUC1). a) RA FLSs were transfected by MUC1 small interfering RNA (siRNA), and the messenger RNA (mRNA) expression levels of proinflammatory cytokines in RA FLSs were evaluated by quantitative real-time polymerase chain reaction (qRT-PCR) (n = 5). b) RA FLSs were treated with GO-203 (5 μM) for 48 hours and expression levels of proinflammatory cytokines were measured (n = 5). Migration of RA FLSs was measured with a Transwell chamber (Corning, USA), and cells seeded onto the upper chamber were allowed to migrate towards medium with 10% fetal bovine serum (FBS) in the lower chamber for six hours. For invasion assay, Transwell chambers coated with Matrigel were used and cells were allowed to invade to the lower chamber filled with 15% FBS for 24 hours. c) to f) Inhibitory effects of MUC1 silencing by siRNA on migration and invasion (c & d; n = 5) in RA FLSs. e) to f) RA FLSs were pretreated with GO-203 (5 μM) for 48 hours and subjected to migration and invasion assay (n = 3). Data are presented as the mean and standard error of the mean (SEM). *p < 0.05, **p < 0.01, independent-samples t-test. CCL-2, C-C motif chemokine ligand-2; Ctr, control; IL, interleukin; NC, negative control; siNC, siRNA of negative control.