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Figure 4


Fig. 4. Time course of TC48 localization in Cos-1 cells. (A) Cos-1 cells transfected with GFP-TC48 were fixed at 12 hours, 24 hours and 48 hours after transfection and stained with anti-giantin mouse monoclonal antibody (red) to stain the Golgi complex, and observed by confocal microscopy. Yellow colour generated in the merged images indicates colocalization of TC48 and Golgi-localized giantin. Notice the progressive increase of GFP-TC48 localization in ER-like reticular structures with increasing time. (B) Cos-1 cells overexpressing GFP-TC48 were fixed after 12 hours of transfection and stained with mouse monoclonal anti-ERGIC-53 antibody. Cy3-conjugated secondary antibodies were used to visualize ERGIC (red). Yellow colour generated in the merged image indicates colocalization of TC48 and ERGIC-localized ERGIC-53. (C) Cos-1 cells were transfected with plasmids expressing VSVG-GFP and HA-TC48. After 15 hours at 40°C, the cells were shifted to 15°C for 3 hours. Cells were fixed and stained with HA-tag antibody (red) and analyzed by confocal microscopy. Bars, 10 µm.





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