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Fig. 8. Steady-state regeneration of Cx26-GFP gap junctions is sensitive to BFA but not nocodazole. NRK cells stably expressing Cx26-GFP were time-lapse imaged following photobleaching of a series of gap junctions (A,D,G, box indicates photobleached area). Cx26-GFP fluorescence steadily recovered within the photobleached area (G, box). ER-to-Golgi transport was disrupted by a 45 minute pre-incubation with BFA followed by FRAP in the presence of BFA (B,E,H, box indicates photobleached area). After 1 hour, there was no detectable Cx26-GFP within the photobleached region of interest (H, box). To disrupt microtubules, nocodazole was added to the cell culture 45 minutes prior to photobleaching of gap junctions in the continued presence of nocodazole (C,F,I). There was significant recovery of Cx26-GFP to the region of interest after 1 hour (I, box), comparable with that occurring under control conditions. Panel inserts represent higher magnifications of the photobleached regions of interest. Experimental sets were repeated seven times with similar results. Bar, 10 µm.
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