Quantification of WT and mutant VWF colocalization with organelle markers. Staining for ER with mouse anti-PDI (ER, 1:100; Abcam) (A), mouse anti-GOLPH4 (cis-Golgi, 1:100; Santa Cruz Biotechnology) (B), goat anti-Rab 27a (pseudo-WPB, 1:50; Santa Cruz Biotechnology) (C), mouse anti-Lamp1 (lysosome, 1:100; R&D Systems) (D), and VWF (rabbit anti-VWF, 1:500; DAKO). Secondary antibodies: Alexa Fluor 488 goat anti-rabbit IgG (Invitrogen), Alexa Fluor 568 donkey anti-goat IgG (Invitrogen), and rhodamine donkey anti-goat IgG (Santa Cruz Biotechnology). 100% indicates homozygous transfections; 50% indicates transfections in the mock heterozygous state where HEK293T cells were transfected with 50% WT vector and 50% mutant vector. Imaging was conducted at room temperature using a Quorum Wave Effects Spinning Disc confocal microscope at ×60 magnification, captured with a Hamamatsu Orca high-resolution camera and ImageJ software analysis. Colocalization of the VWF and the organelles was quantified using MetaMorph microscopy automation and image analysis software (Molecular Devices) to calculate the MCC (N = 40-50 cells per marker; *P < .05; **P < .01).