Drs2-Rcy1 pathway and the v-SNAREs, Snc1 and Snc2, are required for CUPS formation. WT and the indicated deletion or mutant strains expressing Grh1-2xGFP were visualized by epifluorescence microscopy in growth conditions and after incubation in 2% potassium acetate for the indicated times. Cells were classified with normal CUPS (one to three larger foci per cell); intermediate CUPS (int.), where a large focus is observed in addition to smaller structures; and vesiculated CUPS (ves.), where only small foci of Grh1 are observed. Scale bar = 2 μm.