Fig. 6. Colocalization studies of linker-GFP, G2A-GFP and C3S-GFP with the Golgi marker BODIPY-TR-ceramide (A) and cycloheximide treatment of the linker-GFP and C3S-GFP chimeras (B). (A) COS-7 cells were transfected with the linker-GFP (upper panels), G2A-GFP (middle panels) or C3S-GFP (lower panels) constructs and, 36 hours post-transfection, they were incubated with the Golgi apparatus marker BODIPY-Texas Red-ceramide (1.5 µM in DMEM). The GFP fluorescence (left panels) was obtained after excitation at 488 nm whereas the Texas Red fluorescence (middle panels) was obtained after excitation at 543 nm. Right panels show the merge of both fluorescence signals. Bar, 50 µm. (B) Changes induced in the localization of the linker-GFP and C3S-GFP mutants upon treatment with 100 µg/ml cycloheximide for 2 hours. The treatment was performed 24 hours post-transfection.

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