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Fig. 5. Cx43-GFP and Cx26-YFP are transported from the Golgi apparatus in both vesicular- and tubular-like PGCs. Time-lapse images of BICR-M1R_k cells expressing Cx43-GFP (A1,A2) or Cx26-YFP (B1,B2) were obtained from 10 to 40 minutes after the removal of BFA. Movies 1 and 2 (see supplementary material) show that both Cx43-GFP and Cx26-YFP were predominantly in the ER initially (T=10) and were transported to paranuclear locations by 40 minutes (T=40), which reflects the re-organization of the Golgi apparatus. Cells imaged (T=84-90 minutes) after BFA removal revealed a paranuclear Golgi-like distribution of Cx43-GFP (C) and Cx26-YFP (D) with subpopulations of tagged connexins emanating from the Golgi in tubular-like structures (C,D, arrows) and vesicular structures (C,D, arrowheads). Tubular extensions and dynamic vesicle movement are best seen in Movies 3 and 4 (see supplementary material). (E) Cx26-YFP could be observed at the cell surface with a diffused rim appearance (double arrow) in cells imaged 90 minutes following BFA-washout. Movie 5 shows vesicles exiting distal elements of the Golgi apparatus and the appearance of bright clusters at cell surface that move rapidly in the plane of the membrane (see supplementary material). Bars, 10 µm.

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