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Fig. 2. Subcellular localisation of the various constructs and laser confocal microscopy with the Golgi marker ß-cop. COS7 cells were transfected with the various NOS2-GFP constructs and the fluorescence was analysed 36 hours post transfection. The subcellular localisation of the full-length NOS2-GFP chimeras (A) and the (1-94)-NOS2 chimeras (B) was determined. Changes induced in the subcellular distribution of the Myr chimera when treated with 10 µM 2-Br-Palmitate were also determined (C). In order to determine the colocalisation of the Myr, Myr/C3S and A2C NOS2(1-94) chimeras with the Golgi marker ß-cop, double immunofluorescence staining was performed using a Cy3-conjugated secondary antibody. As a positive control, we used residues 1-55 of NOS3 fused to GFP, which is known to become myristoylated and palmitoylated (Navarro-Lérida et al., 2002). Cell nuclei were stained with Hoechst 33258 (blue). Bars, 50 µm.
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