Figure 5.

GBF1 depleted HUVECs produce few but very long VWF structures that remain in the peri-Golgi area

(A) Mean number of VWF “quanta” per cell in control and GBF1-siRNA-treated cells. n = 8 wells, where for each well the mean for each of the 9 fields of view were analyzed, SEM, unpaired t test, p < 0.0001.

(B) Super-resolution structured illumination microscopy (SR-SIM) reconstruction showing control and GBF1-siRNA-treated cells where processed-VWF, green; TGN46, red; and DAPI, blue are shown. Scale bars, 5 μm.

(C) The Golgi area, in control and GBF1-siRNA-treated cells overexpressing GFP-VWF were processed for correlative light electron microscopy (CLEM) analysis. (i) Overlay of confocal and bright field image. Scale bar, 10 μm. (ii) Overlay of confocal and a single EM section. (iii) Inset of region of interest (box in [i] and [ii]), 3D reconstruction of WPBs (mock cell 27 EM slices and siGBF1 cell, 15 EM slices). Each WPB has been pseudo-colored in a different color. Scale bar, 2 μm. (iv) Inset of region of interest, single EM section. Arrowheads point to WPBs. Scale bar, 2 μm.

See also Figure S3.