Fig. 4

c-Raf was involved in MCP-1-mediated osteosarcoma migration. a At different MCP-1 stimulation durations (0, 10, 15, 30, and 60 min), c-Raf phosphorylated protein and c-Raf total protein were measured. b MG63 cells were pretreated with 0.1% DMSO as a control, GW5074 for 30 min and then incubated with MCP-1 for 30 min. c-Raf phosphorylation was examined using Western blotting. c The MG63 cells were treated with GW5074 (5 μM), and the degree of cell migration was measured with MCP-1 stimulation. d and e MMP-9 mRNA and protein expression were detected after the MG63 cells were treated with GW5074 and MCP-1 stimulation. f-g After the control shRNA and c-Raf shRNA were transfected in the MG3 cells and incubated for 24 h, a cell migration assay was performed with or without MCP-1 stimulation. h The MG63 cells were transfected with c-Raf shRNA and incubated for 24 h, and MMP-9 mRNA expression was measured using the qPCR assay. Results are expressed as mean ± SEM, n = 4. *p < 0.05 compared with control or control shRNA groups; #p < 0.05 compared with the MCP-1-treated group