GMAP_N and α-synuclein cause accumulations of distinct vesicular structures in yeast cells. (A, top) Schematic diagram of GMAP_N-mCherry (mCh). (bottom) Localization of this construct in yeast cells. (B) Representative transmission EM images of cells expressing GMAP_N-GFP. (C, top) Schematic diagram of α-synuclein–GFP. (bottom) Localization in yeast cells. (D) Representative EM image of cells expressing α-synuclein–GFP. (E) Immunogold labeling of cells expressing GMAP_N-GFP using antibodies against GFP. Arrows show vesicular structures. N, nucleus; V, vacuole; ER, endoplasmic reticulum; CW, cell wall. (F) Strains IPY4 GMAP_N-GFP and IPY5 α-synuclein (α-syn)–GFP were grown overnight under repressing conditions and then shifted (open symbols) or not shifted (closed symbols) to induction medium for the indicated times, and absorbance (Abs) of the cultures at OD₆₀₀ was monitored. (G) Localization of GMAP_N-mCherry and α-synuclein–GFP expressed together in wild-type yeast cells. (top and bottom) Two optical sections of a z stack obtained for a single cell. BY4742 cells carrying the indicated proteins on plasmids were grown overnight (A–E) or for 4 h (G) under inducing conditions and imaged.