Figure 5. p45 MET fragment promotes cell migration of MDCK epithelial cells.

(A) Cell lysates from MDCK cells stably expressing (Clone 9 and 41) or not (mock) the flagged MET fragment beginning at the C970 amino acid (flag-p45 MET) were analyzed in duplicate by western blotting with an antibody directed against the human MET C-terminal region and GAPDH. Arrowheads indicate phosphorylated MET and the flag-p45 MET fragment. (B) MDCK stably expressing or not flag-p45 MET were seeded at low density. The next day the cells were treated 24h with 10ng/ml of HGF/SF. Cell islets were then fixed and stained (scale bar = 50 μm). (C) To evaluate cell scattering, cell density of more than 150 islets were calculated and shown as reciprocal values in a boxplot (the islet area divided by the number of cells in each islet). p value of student's t test is shown (**p < 0.001). (D) MDCK stably expressing or not flag-p45 MET were stained with fluorescent Dil-C12, seeded at low density and treated with HGF/SF 10ng/ml. Cells were tracked every 10min during 20 h by fluorescent video microscopy. Trajectories of 50 individual cells placed at the same origin were shown. (E) MDCK stably expressing or not flag-p45 MET were seeded at high density on modified Boyden chambers coated with Matrigel with 30 ng/ml of HGF in the bottom compartment. Two days later, cells of the transwell bottom surface were counted (relative number cells to control; n = 3, ±standard deviation).