JCS02569 Supplementary Material

Files in this Data Supplement:

• Movie 1 - Movie 1. Movie submitted with Fig. 6A (Visualization of ER to Golgi transport of Cx43-GFP following removal of BFA). BICR-M1R_k cells incubated for 6 hours in the presence of 5 mg/ml BFA were time-lapse imaged between 10 minutes and 40 minutes after the removal of BFA. Note the movement of a predominantly ER-like pattern of Cx43-GFP to a paranuclear Golgi-like pattern. Movie includes 50 scans representing 30 minutes of real-time imaging.

• Movie 2 - Movie 2. Movie submitted with Fig. 6B (Visualization of ER to Golgi transport of Cx26-YFP following BFA removal). BICR-M1R_k cells incubated for 6 hours in the presence of 5 mg/ml BFA and imaged between 10 minutes and 40 minutes after the removal of BFA. The predominant ER-like pattern of Cx26-YFP redistributes to a Golgi-like distribution by 40 minutes. Movie includes 50 scans representing 30 min of real-time imaging.

• Movie 3 - Movie 3. Movie submitted with Fig. 6C (Tubular extensions and vesicles containing Cx43-GFP exiting the Golgi apparatus). This time-lapse series was acquired from 80 minutes to 125 minutes after the removal of BFA. Tubular extensions are seen exiting the Golgi apparatus as well as numerous vesicles. Movie includes 50 scans representing 30 minutes of real-time imaging.

• Movie 4 - Movie 4. Movie submitted with Fig. 6D. (Visualization of tubular extensions containing Cx26-YFP). A tubular extension containing Cx26-YFP is observed emanating from the Golgi apparatus. Note it extends and a vesicle appears to bud off it. Numerous vesicles are also observed. This movie was acquired 80-110 minutes after BFA removal. Movie includes 50 scans representing 30 minutes of real-time imaging.

• Movie 5 - Movie 5. Movie submitted with Fig. 6E. (Visualization of random transport of Cx26-YFP to the cell surface). Diffuse fluorescence is observed evenly at the cell surface indicating random, undirected transport of PGCs to the cell surface. Bright clusters appear at regions of cell-cell contact which rapidly disappear or move within the plane of the membrane prior to clustering into gap junction plaques. Movie was taken 90 to 120 minutes after BFA removal. Movie includes 50 scans representing 30 minutes of real-time imaging.

• Movie 6 - Movie 6. Movie submitted with Fig. 7I. (Visualization of dynamic vesicular transport of a Golgi-localized Cx26 mutant). The region surrounding the Golgi apparatus of HBL-100 cells expressing D66K-GFP was photobleached and time-lapse imaged to observe the dynamics of any PGCs. Even though this is a Golgi-localized mutant, multiple vesicles are observed emanating from the Golgi apparatus. Movie includes 10 minutes of real-time imaging.

• Movie 7 - Movie 7. Movie submitted with Table 1 (Nocodazole decreases the distance traveled by vesicles containing Cx26-YFP). NRK cells expressing Cx26-YFP were time-lapse imaged prior to the addition of nocodazole. After nocodazole was added, the same cells were time-lapse imaged. Note the decrease in movement of vesicles containing Cx26-YFP. Movie represents 25 scans and 18 minutes of real-time imaging in the absence of nocodazole (left-hand panel) and 18 minutes in the presence of nocodazole (right-hand panel).