Figure 4

MIF activates p44/p42 MAPK pathway and stimulates MIP-2 release from macrophages. A. RAW264.7 cells were stimulated with 100 ng/ml MIF for 5, 10, 15, 30 or 60 min. After stimulation the cells were lysed and boiled for SDS-PAGE analysis. Transferred proteins were detected by anti-phospho and total p44/p42 MAPK antibodies. Data represents four independent experiments. B. RAW264.7 cells were stimulated with 5, 20, 100 ng/ml MIF for 10 min. Proteins were detected by anti-phospho and total p44/p42 MAPK antibodies. Data represents three independent experiments. C. RAW264.7 cells were stimulated with different concentrations of MIF (20, 50, 100 ng/ml) or control media for 18 hours at 37°C. After 18 hours supernatants were collected and MIP-2 concentrations were measured by ELISA. Bars indicate mean ± SD (n = 3/group). Data is representative of four independent experiments. *P < 0.05 compared with control media.