Figure 5.

GFP-golgin-160 and GFP-golgin-160(D59A) localize to the Golgi. Stable HeLa lines expressing either GFP-golgin-160 (GFP fused to the NH₂ terminus of wild-type golgin-160) or GFP-golgin-160(D59A) (GFP fused to the NH₂ terminus of the D59A golgin-160 mutant) were generated. Cells were fixed, permeabilized, double stained with anti-GM130 and anti-GFP, and examined by confocal immunofluorescence microscopy. GM-130 antibodies were visualized with goat anti–mouse IgG and assigned the color red (a and d), whereas GFP antibodies were visualized with fluorescein-conjugated goat anti–rabbit and assigned the color green (b and e). When images were merged, overlapping red and green pixels appeared orange/yellow (c and f). Experiments were repeated on four separate occasions with similar results. Bars, 10 μm.