Figure 5

The effect of NEM on the localization of expressed syntaxin 2 and 3 in NRK cells. Syntaxin 2 and 3 were expressed in NRK cells and stained with syntaxin 2 or 3 antiserum. (A) Control cells show localization of expressed syntaxins both at the plasma membrane and in intracellular sites. The control cells were treated with 50 μg/ml cycloheximide (controls). Also expressed syntaxin 2 and 3 accumulated into intracellular sites in the presence of NEM. (B) NEM-treated cells were double stained using syntaxin 2 or syntaxin 3 anti-serum and monoclonal antibodies against transferrin receptor (Ox26) or TGN38 as in Fig. 4. The yellow colour in merged images reveals the co-localization. Exposure in the pictures was adjusted so that only expressed syntaxins can be seen. Conventional fluorescence images were viewed using an Olympus AX70 fluorescence microscope (A). Confocal fluorescence images were viewed using a Leica SP1 microscope system (B). Bars, 10 μm.