Fig. 3.

Intracellular localization of mVWF.

Wt (A) and mVWF (D) were transiently expressed in HEK293 cells. 48 h after transfection, cells were fixed and VWF proteins (A,D) and PDI (B,E) were detected by indirect immunofluorescence employing rabbit anti-VWF and mouse anti-PDI antibodies, respectively. Z-stacks were recorded with a confocal microscope using an HC PL APO CS2 63.0x1.40 OIL UV objective and the following settings: image size of 512 × 512, laser power of the 543 and 488 lasers was set to 9 % and 20 %, respectively. Overlays are shown in (C,F). 3D reconstruction was performed using the LAS software (Leica). Scale bar represents 25 μm. For movies of the rotating complete 3D reconstruction please refer to supplemental Movies S1 (rwtVWF) and S2 (rmVWF).