Colocalization of FITC-FXII with uPAR or gC1qR on HUVECs.

Nonpermeabilized, paraformaldehyde (2%)–fixed HUVECs were grown on microscope slides. HUVECs were incubated in HCB containing 10 μM ZnCl₂ with 300 nM FITC-FXII in the presence of anti-gC1qR (4 μg/mL) or anti-uPAR (4 μg/mL) antibodies for 1 hour at 37°C. The dual detection of the antigens was performed with a secondary antibody labeled with Alexa Fluor 594–labeled goat antimouse IgG conjugate (10 μg/mL) and FITC-FXII as described in “Materials and methods.” The panels are photomicrographs of the laser scanning confocal microscopy. The figure is a representative presentation of 3 independent experiments.