Fig. 2. GFP-GBF1, like endogenous GBF1, is recruited to cis-Golgi membranes and peripheral VTCs membranes. (A) Schematic representation of GFP-GBF1: EGFP (green) was fused in frame with full length CHO-derived GBF1 (orange) at its N terminus. Additional residues encoded by plasmid-derived and UTR sequences shown in blue. (B) NRK cells stably expressing GFP-GBF1 were fixed and processed for either double-label confocal IF by staining with affinity purified polyclonal antibody against GFP (a) and monoclonal antibody against p115 (3A10) (c) or single-label IF by staining with polyclonal antibody against BIG1 (9D3) (f) or monoclonal antibody against ß-COP (M3A5) (i). Arrowheads mark puncta revealed by GFP fluorescence (d,g) or GFP antibodies (a). Middle panels (b,e,h) show merged left and right images. Insets show threefold magnification of the area indicated by arrow. Bars, 10 µm.