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. 2019 Jun 3;49(5):786–801.e6. doi: 10.1016/j.devcel.2019.04.006

Figure 1.

Figure 1

ARF1 and ARF4 and Their GEF GBF1 Are Involved in WPB Biogenesis

(A) Immunofluorescence images showing HUVECs treated with the indicated siRNAs. Representative images acquired using an Opera confocal and used for image quantification, stained for pro-VWF, green; TGN46, red; and DAPI, blue. Scale bars, 20 μm.

(B) Mean number of WPB per cell in control and ARF siRNA treated cells. n = 5 wells, where for each well the mean for each of the 9 fields of view were analyzed, SEM., one-way ANOVA with Dunnett’s multiple comparisons test, ∗p < 0.5, ∗∗p < 0.05, ∗∗∗p < 0.005, ∗∗∗∗p < 0.0005.

(C) Proportion of secreted VWF from total VWF in control and ARF siRNA treated cells upon 30 min histamine stimulation. Results standardized to the amount secreted by control cells in each experiment. n = 3 independent experiments, SEM., one-way ANOVA with Dunnett’s multiple comparisons test, ∗p < 0.5, ∗∗p < 0.05, ∗∗∗p < 0.005, ∗∗∗∗p < 0.0005.

(D) Mean number of WPB per cell in control and ARF GEF siRNA treated cells. n = 5 wells, where for each well the mean for each of the 9 fields of view were analyzed, SEM., one-way ANOVA with Dunnett’s multiple comparisons test, ∗p < 0.5, ∗∗p < 0.05, ∗∗∗p < 0.005, ∗∗∗∗p < 0.0005.

(E) Proportion of secreted VWF from total VWF in control and ARF GEF siRNA treated cells upon 30 min histamine stimulation. Results standardized to the amount secreted by control cells in each experiment. n = 3 independent experiments, SEM., one-way ANOVA with Dunnett’s multiple comparisons test, ∗p < 0.5, ∗∗p < 0.05, ∗∗∗p < 0.005, ∗∗∗∗p < 0.0005.

(F) Mean number of WPB per cell in control and ARF1 and ARF4 and GBF1 siRNA treated cells. n = 5 wells, where for each well the mean for each of the 9 fields of view were analyzed, SEM., one-way ANOVA with Dunnett’s multiple comparisons test, ∗p < 0.5, ∗∗p < 0.05, ∗∗∗p < 0.005, ∗∗∗∗p < 0.0005.

(G) Proportion of secreted VWF from total VWF in control and ARF1 and ARF4 and GBF1 siRNA treated cells upon 30 min histamine stimulation. Results standardized to the amount secreted by control cells in each experiment. n = 3 independent experiments, SEM., one-way ANOVA with Dunnett’s multiple comparisons test, ∗p < 0.5, ∗∗p < 0.05, ∗∗∗p < 0.005, ∗∗∗∗p < 0.0005.

(H) GBF1 depletion in endothelial cells does not induce gross morphological Golgi defects but causes dispersion of the ERGIC. Immunofluorescence confocal images of control and GBF1-siRNA-treated HUVECs for various ER-Golgi proteins. Scale bars, 10 μm.

(I) Super-resolution structured illumination microscopy (SR-SIM) reconstruction of control and GBF1-siRNA-treated cells stained for TGN46, green and GM130, red. Scale bars, 5 μm.

See also Figures S1 and S2.