Vps1 dynamin family protein localizes to SNX-BAR–coated endosomes. The top row shows diploid cells expressing one untagged copy of Vps1 and one GFP-tagged copy of Vps1 (Vps1-GFP) in cells coexpressing Mvp1-2xRFP (top) or Vps17-2xRFP (middle). Vacuoles (blue) were visualized by staining with the vital dye, CMAC. An expanded view of the boxed region in the “merge” column is shown in the “inset” column. The bottom row shows Vps1-GFP in homozygous diploid mvp1Δ mvp1Δ cells. One deconvolved image of a z-series containing the endosome or vacuole marker is shown. Bar: (main panels) 5 µm; (insets) 2.5 µm. Two methods for representing colocalization of Vps1-GFP with Vps17-2xRFP are depicted in B and C. Refer to text for a detailed description of the quantitation methods. (B) Distribution of the proportion of Vps1-GFP and Vps17-2xRFP double-positive endosomes. Wild-type (dark blue) and mvp1Δ mvp1Δ (light blue) cells (n = 20) were binned according to the proportion of Vps1-GFP and Vps17-2xRFP double-positive endosomes. (C) The proportion of endosomes (wild type: n = 64 of endosomes; mvp1Δ: n = 82 endosomes derived from cells analyzed in B) decorated by both Vps1-GFP and Vps17-2xRFP are shown. Standard deviations are indicated; P < 0.0001 (unpaired t test).