Figure 2From: Macrophage migration inhibitory factor enhances osteoclastogenesis through upregulation of RANKL expression from fibroblast-like synoviocytes in patients with rheumatoid arthritisThe effect of MIF on the expression of RANKL in RA human synovial fibroblasts. (a) Isolated RA synovial fibroblasts were incubated with rhMIF (0.1 to 10 ng/mL) for 72 hours, and mRNA was extracted and measured using real-time PCR. (b) Isolated RA synovial fibroblasts were incubated with rhMIF (0.1 to 10 ng/mL) for 72 hours, and protein was extracted and measured using western blot analysis. (c) RA synovial fibroblasts were cultured with 0.1 to 10 ng/mL of rhMIF for 72 hours and stained with an anti-RANKL antibody (red) (original magnification 400×). (d) Effect of neutralizing agents for known osteoclastogenic factors on MIF-induced RANKL expression. RA synovial fibroblasts were treated with rhMIF 5 ng/mL for 72 hours in the presence or absence of anti-IL-1β, anti-TNF-α, or anti-IL-6. RANKL mRNA expression was quantified using real-time PCR. (e) Isolated RA synovial fibroblasts were incubated with rhMIF (0.1 to 10 ng/mL) for 72 hours, and MIF-induced IL-1β mRNA expression was measured using RT-PCR. The data represent the mean and standard deviation of three separate experiments. *P < 0.05 and **P < 0.005. MIF, macrophage migration inhibitory factor; RA, rheumatoid arthritis; RANKL, receptor activator of nuclear factor kappa-B ligand.Back to article page