The ALPS motif of GMAP-210 acts as a localization determinant to cytoplasmic vesicular structures. (A) Localization of a truncated version of GMAP_N-mCherry with the ALPS motif deleted (schematic diagram, left). Fluorescence (left) and overlay onto a differential interference contrast image (right) are shown. (B, left) Localization (bottom) of constructs is shown in the schematic diagrams (top). ALPS–Smc1-CC was constructed by replacing the CC region of GMAP_N with that of the nuclear protein Smc1p (aa 158–374). (right) For quantifications, 60–130 structures for each GFP fusion protein were scored for localization to the nucleus, as visualized by Hoechst staining, or to the cytoplasm. Results shown are representative of two independent experiments. Outlines of cells are indicated by dashed lines. (C) Representative EM image of cells expressing the ALPS–Smc1-CC–GFP chimeric protein. Inset shows a higher magnification of the boxed region. BY4742 cells expressing the indicated proteins were grown overnight under inducing conditions before imaging.