Figure 3

(a) Real-time RT–PCR to quantify the MET mRNA expression level: **P<0.001 for comparison of OH931 or NMFH-1 myxofibrosarcoma cell line with CCD966SK fibroblasts; ##P<0.001 for comparison between OH931 and NMFH-1 cells. (b) Western blotting assays: when compared with CCD966SK fibroblasts, MET protein overexpression was apparent with activating phosphorylation in OH931 myxofibrosarcoma cells as seen by probing with phospho-METTyr1234/1235, whereas there was only modest expression of total MET protein in NMFH-1 cells without phosphorylation. (c) Kinetics of tyrosine phosphorylation of MET in NMFH-1 myxofibrosarcoma cells in response to exogenous HGF treatment. Cells grown in 6-cm dishes were serum starved overnight, left unstimulated or stimulated with 50 ng/ml HGF for indicated time periods, and then probed with indicated antibodies.