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Fig. 6. Golgi-apparatus-localized Cx26 mutants actively exit the Golgi in the form of dynamic tubular extensions and vesicles. HBL-100 cells stably expressing D66K-GFP (A,C, green) or D66H-GFP (D,F, green) were labeled for the cis-Golgi resident protein, GPP130 (B,E,C,F; red). Both D66H-GFP and D66K-GFP exhibit a high degree of co-localization with the Golgi apparatus (C,F, note yellow color). Cells were counterstained with Hoechst 33342 to denote the nuclei (C,F, blue). HBL-100 cells expressing D66H-GFP were time-lapse imaged following photobleaching of the Golgi population (series G, box indicates photobleached region). A new population of D66H-GFP migrated to the photobleached Golgi apparatus with a 40% recovery of fluorescence in 1 hour (based on the pre-bleached area being 100%). Tubular extensions were observed exiting the Golgi apparatus of HBL-100 cells expressing D66H-GFP (series H, arrows). In series I, the area surrounding the Golgi pool of D66K-GFP mutant was photobleached and time-lapse imaged. Highly dynamic vesicles were observed to emanate from the Golgi apparatus within 15 minutes (series I, arrows and supplementary material Movie 6). Bars, 10 µm.





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