Figure 5
Colocalization of VWF and PDI. wtVWF (A) or VWF mutants p.Cys2771Arg (B) and p.Cys2773Alafs*76 (C) were transiently expressed in HEK293 cells. Forty-eight hours after transfection, cells were fixed and VWF proteins and PDI were detected by indirect immunofluorescence employing rabbit anti-VWF and mouse anti-PDI antibodies; secondary antibodies were goat anti-rabbitAF488 and goat anti-mouseAF546. Images were recorded with a confocal microscope (TCS SP5, Leica), using an HC PL APO CS2 63.0 × 1.40 oil ultraviolet objective and the following settings: image size of 512 × 512 pixels, laser power of the 543 and 488 lasers was set to 9% and 20%, respectively. Colocalization appears yellow in the overlay images; for colocalization analysis, please refer to supplemental Table 1. Scale bars represent 10 µm.

Colocalization of VWF and PDI. wtVWF (A) or VWF mutants p.Cys2771Arg (B) and p.Cys2773Alafs*76 (C) were transiently expressed in HEK293 cells. Forty-eight hours after transfection, cells were fixed and VWF proteins and PDI were detected by indirect immunofluorescence employing rabbit anti-VWF and mouse anti-PDI antibodies; secondary antibodies were goat anti-rabbitAF488 and goat anti-mouseAF546. Images were recorded with a confocal microscope (TCS SP5, Leica), using an HC PL APO CS2 63.0 × 1.40 oil ultraviolet objective and the following settings: image size of 512 × 512 pixels, laser power of the 543 and 488 lasers was set to 9% and 20%, respectively. Colocalization appears yellow in the overlay images; for colocalization analysis, please refer to supplemental Table 1. Scale bars represent 10 µm.

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