Extended Data Fig. 5: Morphology and identity of coated vesicles.
From: In situ cryo-ET defines the ultrastructure of ER exit sites in human cells
a. Gallery of xy slices through COPII-coated vesicles from cryo-lamellae. Top-left panel shows an in vitro reconstituted vesicle for comparison16. b. Gallery of xy slices through COPI-coated vesicles from cryo-lamellae. Top-left panel shows an in vitro reconstituted vesicle for comparison28. c. Gallery of xy slices through other classes of membranes, as described below each panel. d. Slice through the xy plane of a tomogram overlayed with the outline of a mask used to run template matching. The mask at this plane is seen around a COPII-coated vesicle. e. Slice through the xy plane of a tomogram overlayed with the outline of a mask used to run template matching. The mask at this plane is seen around a COPI-coated vesicle. f. Slice through the xy plane of a tomogram overlayed with the outline of a mask used to run template matching. The mask at this plane was randomly applied as control. g. Left: a zoomed in view of a COPII vesicle from a. Right: a map of in vitro assembled COPII inner coat (EMD-19417) was used as template to search cryo-tomograms that were loosely masked around all coated membranes. Highest cross-correlation peaks are mapped onto the tomogram using ArtiaX, and a representative COPII-coated vesicles is shown. Peak positions are shown in green, and their orientations by blue, yellow and red arrows. The mask profile is in white against a black background. h. Left: a zoomed in view of a COPII vesicle from b. Right: a map of in vitro assembled COPI coat (EMD-3968) was used as template to search cryo-tomograms that were loosely masked around all coated membranes. Highest cross-correlation peaks are mapped onto the tomogram using ArtiaX, and a representative COPI-coated vesicles is shown. Peak positions are shown in purple, and their orientations by blue, yellow and red arrows. i. Left: a zoomed in view of a random region from c. Right: a map of in vitro assembled COPI coat (EMD-3968) was used as template to search cryo-tomograms that were loosely masked at random places (a mask was applied to the wrong tomogram). Highest cross-correlation peaks are mapped onto the tomogram using ArtiaX, and a representative COPI-coated vesicles is shown. Peak positions are shown in brown, and their orientations by blue, yellow and red arrows. Scale bars: 100 nm.
