Figure 7

Time course studies of uPAR before and after lipid raft disruption in MDA-MB-231 cells. MDA-MB-231 cells were left untreated or pretreated with 7.5 mM MβCD for one hour at 37°C; washed; fixed with 1% paraformaldehyde at 1, 8 and 24 hrs; permeabilized; incubated with primary antibodies against uPAR and LAMP-1 overnight at 4°C; washed; incubated with appropriate Alexa Fluor secondary antibodies; and mounted with an anti-fading agent containing DAPI. The cells were imaged under an Olympus fluoview confocal microscope.