Skip to main content

Advertisement

Fig. 6 | Arthritis Research & Therapy

Fig. 6

From: CX3CL1 promotes MMP-3 production via the CX3CR1, c-Raf, MEK, ERK, and NF-κB signaling pathway in osteoarthritis synovial fibroblasts

Fig. 6

CX3CL1 induced NF-κB activation through the CX3CR1/c-Raf/MEK/ERK pathway. ac OASFs were incubated with various concentrations of CX3CL1 or pretreated with c-Raf inhibitors (GW5074) or MEK/ERK inhibitors (U0126 and PD98059) for 30 min or transfected with c-Raf shRNA, MEK, ERK, IKKα, and IKKβ mutants before exposure to CX3CL1. NF-κB luciferase activity measured, and results normalized to the β-galactosidase activity. d OASFs were pretreated with c-Raf inhibitors (GW5074) or MEK/ERK inhibitors (U0126 and PD98059) for 30 min followed by stimulation with CX3CL1 for 60 min. p-p65 expression examined using western blot analysis. e Cells were pretreated with 0.1% dimethyl sulfoxide as control, c-Raf inhibitors (GW5074), or MEK/ERK inhibitors (U0126 and PD98059) for 30 min, followed by CX3CL1 treatment for 120 min. ChIP performed using an antibody against p65. One percent of immunoprecipitated chromatin was assayed to verify equal loading (input). fh Protein and mRNA levels of CX3CL1 and MMP-3 in control-shRNA and CX3CL1-shRNA OASFs examined using western blotting and qPCR. Results expressed as mean ± SEM (n = 4). *p < 0.05 compared with control; #p < 0.05 compared with CX3CL1-treated group. MMP matrix metalloproteinase, NF-κB nuclear factor kappa B, sh short hairpin

Back to article page