Figure 2.

GBF1 Depletion Causes an Accumulation of Unprocessed VWF in Endothelial Cells

(A) Amount of VWF mRNA (relative to mock cells). n = 5 independent experiments, SEM, unpaired t test, p = 0.002.

(B) Total amount of VWF protein measured by western blot (WB) (relative to mock cells). n = 4 independent experiments, SEM, unpaired t test, n.s. = not significant.

(C) Representative western blot showing GBF1 protein depletion and VWF processing. “pro-VWF,” processed VWF and “unp-VWF,” unprocessed VWF.

(D) Percentage of pro-VWF from total VWF (quantification from WB). n = 4 independent experiments, SEM, unpaired t test, p < 0.0001.

(E) Immunofluorescence images of HUVECs stained with antibodies targeting unp-VWF, green and pro-VWF, red. Scale bars, 10 μm.

(F) Immunofluorescence images of HUVECs stained for the ER-transmembrane protein calnexin, green and unp-VWF, red. Scale bars, 10 μm.

(G) Immunofluorescence images of HUVECs stained for the ER-luminal protein calreticulin, green and unp-VWF, red. Scale bars, 10 μm.

(H) Immunofluorescence images of HUVECs stained for the ER-luminal protein PDI, green and unp-VWF, red. Scale bars, 10 μm.

See also Figure S3.