GMAP_N and α-synuclein structures colocalize with distinct compartment markers in yeast. (A) Localization in SEY6210 cells of GMAP_N-mCherry with either GFP-Emp47p or pro–α-factor–citrine and of Anp1p–monomeric RFP (mRFP) with GMAP_N-GFP. (top) Representative images of colocalization of GMAP_N-mCherry and GFP-Emp47p. (bottom) For quantifications, between 30 and 100 GMAP_N structures were scored for the presence or absence of the cargo protein, and percentages were calculated (see Materials and methods). (B) Localization in SEY6210 cells of GMAP_N-mCherry with the indicated GFP-tagged protein. Quantifications were performed as in A. (C) Localization in BY4742 of GMAP_N-GFP or α-synuclein–GFP (α-syn) with FM4-64. (D–F) Localization in BY4742 of GMAP_N-mCherry or α-synuclein–mCherry with GFP-tagged charge probes carrying the indicated number of lysine residues. Quantifications (D) and representative images (E and F) are shown. Cells were grown overnight at 23°C (A–C) or 16°C (D–F) under inducing conditions, and then, live cells were imaged and quantified. Means and SDs of at least three independent experiments are shown. Outlines of cells are indicated by dashed lines. Bars, 2 µm.