Colocalization of CK1 or uPAR and vWF antigen on HUVECs.

HUVECs grown on microscope slides were fixed with 2% paraformaldehyde and not permeabilized. The panels to this figure are photomicrographs of the laser scanning confocal micrographs. The cells in this experiment were doubly labeled with the primary and secondary antibodies. The upper left and right panels are nonpermeabilized endothelial cells treated with a rabbit antihuman vWF antibody followed by a second antibody conjugated with FITC. The middle left panel shows endothelial cells incubated with the anti-CK1 antibody anti-GPV20 and the middle right panel shows endothelial cells incubated with an anti-uPAR antibody. Both antibodies were detected with a second antibody conjugated with Alexa Fluor. The lower left panel is combined anti-vWF and anti-CK1 antibodies. The lower right panel is exposure of combined anti-vWF and anti-uPAR antibodies. Recognition of the antibody binding was performed with the secondary antibodies labeled with Alexa Fluor and FITC simultaneously. The figure is a representative presentation of 2 independent experiments.