Co-localization of Met and CD44v6 in internal compartments after HGF stimulation.

<p>(<b>A</b>) Confocal images of endogenous Met and CD44v6 at different time points after HGF-induction. HeLa cells were serum starved, treated for 2 hours with 50 µM cycloheximide, incubated at 4°C for 1 hour with HGF (25 ng/ml) (cold start) and then shifted to 37°C for the indicated times. After fixation cells were stained for Met (red, AF276) and CD44v6 (green, VFF18) and nuclei (blue, Draq5) and imaged with the confocal microscope (LSM510) with a 63× objective (Material and Methods). For each time point, the mean Pearson Coefficient of 4–6 images was calculated using the Imaris software. Scale bar = 15 µm (<b>B</b>) Intensity profiles of the emission wavelengths of the Met and CD44v6 signals along lines drawn across the plasma membrane or vesicles containing both Met and CD44v6. The intensity of the respective wavelengths is plotted against the distance in µm. (<b>C</b>) Co-immunoprecipitation of CD44v6 and Met upon HGF induction. Serum-starved HeLa cells were induced with HGF (50 ng/ml) for the indicated time periods and Met was immunoprecipitated. The precipitate was subjected to Western Blot analysis and the membrane was blotted for Met, phospho-Met and CD44v6. The experiment was repeated at least three times.