Figure 6.

HGF affects the levels of expression of u-PA and the localization of u-PAR in pancreas cells. Cells were serum starved for 24 h and treated with HGF (20 ng/ml) for the time periods shown, in the absence or in the presence of amiloride (Am) (0.2 mmol/L) or EACA (50 mmol/L), conditions not permissive for scatter. A: Northern blotting with total RNA (15 μg) from NPCs and IMIM-PC-2 pancreas cancer cells was hybridized to u-PA cDNA probe and subsequently to a thymosin β4 cDNA probe to normalize for RNA deposits. Filters were exposed for 48 h to autoradiography. B: Equal amounts of protein (50 μg) from cell lysates were analyzed by Western blotting with anti-u-PA antibody. C: Twenty-four hour conditioned medium (20 μl) from the same experiment shown in B was analyzed by gelatin zymography in the presence of plasminogen as a substrate. D: Equal amounts of 1% Triton X-100-soluble proteins (s) (50 μg) and equivalent amounts of the insoluble fractions (i) were analyzed by Western blotting using anti-u-PAR antibody. Mobility markers correspond to ovalbumin (53 kd) and carbonic anhydrase (35 kd).