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Fig. 7. NHE7 and SCAMPs colocalize with syntaxin 6 and transferrin receptor. Endogenous SCAMP1, -2, and -5 were visualized in PC12 cells using anti-SCAMP polyclonal rabbit antibodies. To determine intracellular localization of NHE7, PC12 cells were transiently transfected with myc-tagged NHE7 and an anti-myc polyclonal rabbit antibody was utilized. Anti-syntaxin 6 (TGN marker) and transferrin receptor (recycling endosomal marker) monoclonal mouse antibodies were used as organellar markers. NHE7 or SCAMPs were visualized with Alexa 488-conjugated goat anti-rabbit IgG (green) and syntaxin 6 or transferrin receptor was visualized with Alexa 568-conjugated anti-mouse IgG (red). NHE7 exhibited the best colocalization with syntaxin 6, whereas partial colocalization with transferrin receptor was also observed. SCAMPs showed close association with both syntaxin 6 and transferrin receptor, suggesting a more widespread distribution.





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