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Figure 5


Fig. 5. Cytoskeletal rearrangements in HEK-293/uPAR-25 cells exposed to GFDp and/or CPp. Cells were collected by a mild tryspinization, incubated in suspension with diluents or His-uPA or GFDp or CPp at the indicated concentrations for 1 hour at 23°C. In the combinations, a molar ratio of 1:1 CPp:GFDp was employed. When specified, cells were incubated with diluents (none), or preloaded with 50 µg/ml RGD peptide, 5 µg/ml anti-uPAR 399 polyclonal or with the specific anti-integrin antibodies at a dilution of 1:30 for 1 hour. F-actin was detected with Rhodamine-phalloidin; uPAR was detected with anti-uPAR 399 polyclonal antibody followed by a secondary FITC-conjugated anti-rabbit antibody, as specified in Materials and Methods. (A) A representative confocal image of GFDp-treated HEK-293/uPAR-25 cells double-stained with Rhodamine-phalloidin and anti-uPAR antibodies. Original magnification, x630. (B,C) Values reported on the y-axis correspond to the net percentage of cells exhibiting F-actin-enriched protrusions upon exposure to the indicated effectors. Data represent the mean of three independent experiments ± s.d. (error bars), performed in triplicate and evaluated by two independent observers. *P<0.005; **P<0.0001.





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