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. 2017 Jun 5;216(6):1745–1759. doi: 10.1083/jcb.201702135

Figure 2.

Figure 2.

Large COPII vesicles are hollow membranous containers. 3D STORM images of large SEC31A (A) or KLHL12-FLAG (B) puncta marked as red arrowheads in KI6 cells after 7.5-h induction. Position in depth (z axis) is represented by color according to the color bar in Ai and Bi. (i) Overview of an area of the cell imaged by STORM: SEC31A (A) or FLAG (B). (ii) Magnified maximum xy projection of a large SEC31A (A) or FLAG (B) structure. Virtual cross sections of this structure in XZ (iii) and YZ (iv) planes reveal a hollow compartment encapsulated by SEC31A (A) or KLHL12-FLAG (B). (C) CLEM-resolved overlapping immunofluorescent PC1 (green; i) and SEC31A (red; ii) puncta as single membrane-bound compartments in cultured Saos-2 cells expressing endogenous levels of PC1 and KLHL12. (iii) Representative cell showing colocalizing puncta (n = 36; an example was indicated by white arrowhead) was processed for CLEM. Image shows overlay of the merged fluorescent channels (green and red) on a thin-section (70-nm) TEM image (Z1-11). Serial sections of 70-nm thicknesses through structure of interest. (iv) Magnified view of lipid bilayer from an area of the single membrane enclosing the organelle. n = 2. Bars: (Ai) 1 µm; (Bi) 1 µm; (Aii–Aiv) 200 nm; (Bii–Biv) 200 nm; (Ci–Ciii) 1 µm; (Z1–Z11) 100 nm; (Civ) 10 nm.