Localization of CK1 and gC1qR and vWF and gC1qR 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 figure were doubly labeled with primary and secondary antibodies. The panels on the left are cells that were doubly labeled with anti-gC1qR and anti-CK1 (anti-GPV20). The panels on the right are cells that were doubly labeled with anti-vWF and anti-gC1qR. Upper left panel represents cells that have been labeled with mouse anti-gC1qR (4 μg/mL); upper right panel, rabbit antisera to human vWF. Their detection was performed with a second antibody conjugated with FITC. Middle left panel represents cells that have been labeled with goat anti-GPV20 (200 μg/mL) (anti-CK1 antibody); middle right panel, mouse anti-gC1qR (4 μg/mL). Their detection was performed with secondary antibodies conjugated with Alexa Fluor. Lower left panel represents detection of the 2 labels on these cells that were treated with both goat anti-GPV20 (200 μg/mL) and mouse anti-gC1qR (4 μg/mL); lower right panel is a mixture of rabbit antihuman vWF antisera and mouse anti-gC1qR (4 μg/mL). 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 3 independent experiments.