Fig. 6. Loss of Vps20p or Vta1p causes defects in Ste3p localisation to the vacuole. Wild-type (SF838-9D), vps20 (SF838-9Dvpl10) and vta1 (PLY3046) cells were transformed with a low-copy plasmid encoding Ste3-GFP in combination with an empty URA3 containing centromeric plasmid or centromeric plasmid containing the wild-type VTA1 or VPS20 gene as indicated. The localisation of Ste3-GFP was then assessed by fluorescence microscopy together with DIC imaging to identify yeast vacuoles. Cells were resuspended in 1% sodium azide, 1% sodium fluoride, 100 mM Tris pH 8.0 prior to fluorescence and DIC microscopy. Bar, 5 µm.