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Figure 8


Fig. 8. PAI1-mediated disruption of focal adhesions does not require uPAR. (A) MG-63 cells were plated overnight in complete medium. Cells were subsequently fixed, permeabilized and stained with uPAR polyclonal antibody (panel a) and 15F11 monoclonal antibody against β5 integrin (panel b). uPAR and β5 integrin were visualized by indirect immunofluorescence. Panel c shows the merged image. Scale bar, 20 µm. (B) Cells stained for uPAR and paxillin. Panels a and b show uPAR and paxillin staining in cells transfected with control siRNA. Panels d and e show uPAR and paxillin staining in uPAR knockdown cells. Panels c and f are the merged images. Yellow indicates colocalization of uPAR and β5 integrin. Scale bar, 20 µm. (C) Cells transfected with uPAR siRNA were plated overnight in complete medium and then incubated for 3 hours in DMEM in the presence or absence of PAI1. Scale bar, 20 µm. Cells were subsequently fixed, permeabilized and stained. uPAR and β5 integrin were visualized by indirect immunofluorescence in untreated cells (panels a and b) and PAI1 treated cells (panels c and d).





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