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. 2014 Mar 18;289(18):12791–12804. doi: 10.1074/jbc.M113.498576

FIGURE 5.

FIGURE 5.

uPAR ligation rescues the integrin activation and migration defect upon lipid raft disruption by CD. Lipid rafts were disrupted by cholesterol depletion using treatment with or without CD (10 mm, 30 min) followed by treatment with or without CD and with or without ATF (10 nm, 60 min). A, validation of the lipid raft isolation procedure. Cells were treated with or without CD followed by Western blot for caveolin-1 (raft marker) or transferrin receptor (TsfR) (non-raft marker). B and C, filipin staining of cholesterol. B, pseudocolored photomicrographs of filipin staining (red/yellow, high; blue/green, low). C, quantification of plasma membrane/perinuclear area ratio of filipin staining. *, p < 0.05 comparing conditions with or without CD. D, effect of CD on the integrin activation/FN-coated bead binding assay as above. +, p < 0.05 comparing values with or without CD under basal (−uPA) conditions. E, monolayer migration assay as above. +, p < 0.05 comparing values with or without CD under basal (−uPA) conditions. Error bars, S.E.