Figure 1.

Differential trafficking features of recycling cargoes. (A) WT cells expressing Snc1 and Snc2 tagged with either GFP or a fusion of GFP with the catalytic domain of DUb UL36 (DUb + GFP) expressed from the CUP1 promoter were imaged by Airyscan microscopy. (B) Mup1 and Fur4 expressed from their endogenous promoters and fused to C-terminal GFP or GFP-DUb tags were imaged by Airyscan microscopy. Where indicated, 20 µg/ml methionine (+Met) and 40 µg/ml uracil (+Ura) were added to media 1 h prior to imaging. (C) Time-lapse microscopy of cells expressing GFP-Snc2 (left) or Fur4-mNG (right). (D and E) WT cells expressing fluorescently labeled Snc1, Mup1, and Fur4 were imaged, with example cell cycle stages depicted, and fluorescence in mother–daughter pairs quantified. *, P < 0.002. (F–J). Indicated GFP tagged cargoes were imaged by Airyscan microscopy in Sec7-mCherry (upper micrographs) or Vps4-mCherry (lower micrographs) cells, with associated jitter plots of Mander’s overlap coefficients (MOC). *, P < 0.0001 from unpaired t test. (K) Schematic summarizing distinct yeast endosomal recycling pathways. Scale bar, 5 µm.