Figure 3
Figure 3. Electron tomography reveals numerous connections between the Golgi apparatus and WPBs found after 4 hours of transfection. Example of a WPB 4 hours after transfection displaying numerous connections to the Golgi when analyzed by electron tomography. The WPB was localized using CLEM. (A) Two-dimensional image of an immature WPB that was observed in close proximity to the Golgi. (B-C) Tomogram sections showing the connections (arrows) to the Golgi. Also note that the membrane that is not associating with the Golgi is coated by clathrin (arrowheads). (D) A 3-dimensional model of the forming WPB (green) showing the numerous connections (arrows). (E) The addition of a 3-dimensional representation of the Golgi (light yellow) reveals its complex organization around the forming WPB (green). See also supplemental Video 1. Scale bars: (A-B) 250 nm. Electron micrographs and tilt series for tomography were acquired using an FEI Tecnai 12 at 120 kV and using an FEI Eagle 4k × 4k CCD camera. Tilt series for tomography were collected with SerialEM. Tomogram reconstructions were performed in IMOD. Segmentation was performed in Amira (Visualization Sciences Group, FEI). Go, Golgi.

Electron tomography reveals numerous connections between the Golgi apparatus and WPBs found after 4 hours of transfection. Example of a WPB 4 hours after transfection displaying numerous connections to the Golgi when analyzed by electron tomography. The WPB was localized using CLEM. (A) Two-dimensional image of an immature WPB that was observed in close proximity to the Golgi. (B-C) Tomogram sections showing the connections (arrows) to the Golgi. Also note that the membrane that is not associating with the Golgi is coated by clathrin (arrowheads). (D) A 3-dimensional model of the forming WPB (green) showing the numerous connections (arrows). (E) The addition of a 3-dimensional representation of the Golgi (light yellow) reveals its complex organization around the forming WPB (green). See also supplemental Video 1. Scale bars: (A-B) 250 nm. Electron micrographs and tilt series for tomography were acquired using an FEI Tecnai 12 at 120 kV and using an FEI Eagle 4k × 4k CCD camera. Tilt series for tomography were collected with SerialEM. Tomogram reconstructions were performed in IMOD. Segmentation was performed in Amira (Visualization Sciences Group, FEI). Go, Golgi.

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