Figure S5. MDCK cell morphogenesis assay and signalling pathway activation using K1K1, K1K1S2, and K1K1S4 mutants.

(A) MDCK cell (∼500) were seeded into a thick layer of a 1:1 collagen/Growth Factor Reduced Matrigel and treated with semi-log dilutions (100 pM to 300 nM) of hepatocyte growth factor/scatter factor, K1K1, K1K1S2, and K1K1S4 twice a week for 1 mo. The cells were fixed and stained with Evans blue and DAPI. The image of the full plate is presented. (B) Colonies were observed in bright field using inverted microscope observed 4× objective on a Nikon Eclipse TS100 microscope. (C) Phosphorylation analysis of MET signalling pathway by Western blot on HeLa cell lysates after stimulation with semi-log dilution (0.3–30 nM) of K1K1, K1K1S2, or K1KS4 for 10 min. Loading controls are based on total MET, total Akt, and total ERK present in each lane.

Source data are available for this figure.