Fig. 1. Expression of full-length GCC185 results in dispersal of GCC185-labelled structures. (A) COS cells were transfected with myc-GCC185, fixed, permeabilized and stained with anti-myc monoclonal antibodies and FITC-anti-mouse Ig. Shown are three images expressing different levels of GCC185. (B) COS cells were co-transfected with myc-GCC185 and GFP-GCC185_GRIP constructs, fixed, permeabilized and stained with anti-myc monoclonal antibody and Alexa 568 anti-mouse IgG. Superimposed images (merge) reveal regions of colocalization (arrows). (C) COS cells were co-transfected with myc-GCC185 and GFP-golgin97_GRIP constructs, fixed, permeabilized and stained with anti-myc monoclonal antibody and Alexa 568 anti-mouse IgG. (D) COS cells were co-transfected with constructs encoding GFP-GCC185_GRIP and myc-GCC185. Total extracts were prepared after 48 hours transfection as described in Materials and Methods, and lysates were either immunoprecipitated (IP) with anti-GFP antibody (GFP), anti-myc monoclonal antibody (Myc) or with an irrelevant monoclonal antibody (Control). Immune complexes were collected and subjected to SDS-PAGE under reducing conditions. After transfer to PVDF membranes, membranes were immunoblotted with anti-GFP antibody or anti-myc antibody as indicated using a chemiluminescence detection system. Bars: 10 µm.