Fig. 3

MCP-1-induced osteosarcoma migration required the CCR2 receptor. a CCR2 and CCR4 inhibitors (400 nM) were added to the cells. After MCP-1 stimulation, MG63 migration was measured. b and c MMP-9 mRNA and protein expression were detected, respectively, with CCR2 and CCR4 inhibitor treatment and MCP-1 stimulation. (D and E) After the MG63 cells were treated with CCR2 mAb for 30 min, cell migration and MMP-9 mRNA expression were measured, respectively. f-h The MG63 cells were transfected with CCR2 siRNA for 24 h, and the CCR2 protein expression, cell migration and MMP-9 mRNA expression were measured with or without MCP-1 stimulation. i CCR2 protein production was detected in the osteoblast cell line hFOB 1.19 and the osteosarcoma cell lines MG63, U2OS and HOS were assessed using Western blotting. j CCR2 protein production was detected in the MG63 cells with different migratory abilities (M10, M20, and M30) through Western blotting. Results are expressed as mean ± SEM, n = 4. *p < 0.05 compared with control, IgG, control siRNA, hFOB1.19 or MG63 groups, respectively; #p < 0.05 compared with the MCP-1-treated group