Journal: BMC Cell Biology

Article Title: Localization of plasma membrane t-SNAREs syntaxin 2 and 3 in intracellular compartments

doi: 10.1186/1471-2121-6-26

Figure Lengend Snippet: Syntaxin 2 localized in perinuclear membrane vesicles and syntaxin 3 localized in the TGN in NEM treated NRK cells . The NRK cells were incubated in the presence of 1 mM NEM for 15 minutes and then further incubated for two hours. Both syntaxin 2 and 3 accumulated into intracellular compartments in the presence of NEM (A,B). The cells were double stained with syntaxin 2 (A) or syntaxin 3 (B) anti-serum and LRSC-conjugated goat anti-rabbit IgG as well as antibodies against transferrin receptor (Ox26) (C) and TGN38 (D) mouse monoclonal antibodies and FITC-conjugated goat anti-mouse IgG. The yellow colour in merged images (E and F) reveals the co-localization. Confocal fluorescence images were viewed using a Leica SP1 microscope system. Bars,10 μm.

Article Snippet: Mouse monoclonal anti-rat TGN38 antibody was a gift from Dr. G. Banting (University of Bristol, Bristol, UK.) and mouse monoclonal anti-rat transferrin receptor Ox26 hybridoma cell line was obtained from Peninsula Laboratories, Inc. (San Carlos, CA).

Techniques: Membrane, Incubation, Staining, Bioprocessing, Fluorescence, Microscopy

Journal: BMC Cell Biology

Article Title: Localization of plasma membrane t-SNAREs syntaxin 2 and 3 in intracellular compartments

doi: 10.1186/1471-2121-6-26

Figure Lengend Snippet: 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.

Article Snippet: Mouse monoclonal anti-rat TGN38 antibody was a gift from Dr. G. Banting (University of Bristol, Bristol, UK.) and mouse monoclonal anti-rat transferrin receptor Ox26 hybridoma cell line was obtained from Peninsula Laboratories, Inc. (San Carlos, CA).

Techniques: Staining, Control, Clinical Proteomics, Membrane, Bioprocessing, Fluorescence, Microscopy