Figure 4.
Ist1 is required for endosomal recycling in yeast. (A) WT cells coexpressing fluorescently labeled versions of Ist1, Vps4, and Sec7 were imaged by Airyscan microscopy. Insets show variation of colocalization. (B) Pearson’s correlation coefficient’s calculated For Ist1-GFP with Sec7-mCherry (gray) and Vps4-mCherry (green). *, P < 0.002 from unpaired t test; individual values are included as jitter plots over histograms and represent n = 27–72 cells per condition. (C) Venn diagram comparing 89 recycling factors (green) with known physical interactors of Vps4 (pink). (D) Localization of stably integrated Ste3-GFP-DUb in indicated strains by Airyscan microscopy. (E) Histogram showing relative growth of rcy1∆ and ist1∆ mutants compared with WT cells across media containing indicated concentrations of tryptophan. *, P < 0.02 from unpaired t test, n = 3. (F) FM4-64 efflux measurements from WT, rcy1∆, and ist1∆ cells loaded with dye for 8 min at RT followed by three times ice-cold media washes. (G) Airyscan microscopy images of cells co-expressing Mup1-GFP and Sec7-mCherry (left) or Vps4-mCherry (right) in WT or ist1∆ cells. Scale bars, 5 µm (white); 0.5 µm (yellow).

Ist1 is required for endosomal recycling in yeast. (A) WT cells coexpressing fluorescently labeled versions of Ist1, Vps4, and Sec7 were imaged by Airyscan microscopy. Insets show variation of colocalization. (B) Pearson’s correlation coefficient’s calculated For Ist1-GFP with Sec7-mCherry (gray) and Vps4-mCherry (green). *, P < 0.002 from unpaired t test; individual values are included as jitter plots over histograms and represent n = 27–72 cells per condition. (C) Venn diagram comparing 89 recycling factors (green) with known physical interactors of Vps4 (pink). (D) Localization of stably integrated Ste3-GFP-DUb in indicated strains by Airyscan microscopy. (E) Histogram showing relative growth of rcy1∆ and ist1∆ mutants compared with WT cells across media containing indicated concentrations of tryptophan. *, P < 0.02 from unpaired t test, n = 3. (F) FM4-64 efflux measurements from WT, rcy1∆, and ist1∆ cells loaded with dye for 8 min at RT followed by three times ice-cold media washes. (G) Airyscan microscopy images of cells co-expressing Mup1-GFP and Sec7-mCherry (left) or Vps4-mCherry (right) in WT or ist1∆ cells. Scale bars, 5 µm (white); 0.5 µm (yellow).

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