Figure 7.

Gsa12 localizes to the vacuole membrane. Wild-type (PHT12) cells expressing both BFP-SKL and GFP-Gsa12 were grown in methanol medium in the presence of FM 4-64 and then adapted to glucose medium for 0, 2, or 4 h as indicated. The cells were visualized in situ by fluorescence microscopy. Peroxisomes were identified by the presence of BFP, which was targeted by its SKL signal, and the vacuole by FM 4-64. GFP-Gsa12 was found in the cytosol and at the vacuole membrane. Colocalization of GFP-Gsa12 with the FM 4-64 marker can be visualized by yellow-orange in the overlayed image. GFP-Gsa12 was also found associated with those vacuole arms that were sequestering the peroxisomes.