Fig. 2.

Mutant VWF is retained in the ER. a HEK293 cells were transfected with plasmids expressing WT or mutant VWF and then stained for VWF (green channel, left panel). PDsRed2-ER Vector was also co-transfected into cells to delineate the endoplasmic reticulum (middle panel, red). The scale bar is 10 μm. The right panel shows the merged confocal images of the first two panels. b Bar graph represents VWF and the ER overlapping as a percentage of total VWF in cells (n = 3 independent experiments). All values are mean ± SD. ***p < 0.001 compared to WT (one-way ANOVA). c After HEK293 cells were transiently transfected with WT or mutant full-length VWF, cell lysates were reduced and analyzed by Western blotting. The empty vector control is shown in lane 1. The bands of pro-VWF are predominant in the lysates of VWF mutant variants (lanes 2–5). Lane 6 shows both pro-VWF and mature VWF in the lysate of WT, and lane 7 shows only mature VWF in normal plasma