Figure 4. Deletion of VPS27 reduces LegC7 toxicity.

(A) BY4742 or vps27∆ strains harboring either the control or LEGC7 ⁺ plasmids were spotted onto CSM-Ura plates containing 2% glucose or 2% galactose in 10-fold serial dilutions (starting OD₆₀₀ = 1.0) and grown at 30°C for 96h. (B) BY4742 or vps27∆ strains expressing GFP or GFP-LegC7 were grown in selective media supplemented with 2% glucose at 30°C, stained with FM4–64, and visualized for GFP and FM4–64 fluorescence. (C) Yeast vps27∆ strains expressing either GFP-CPS or Sna3-GFP harboring the LEGC7 ⁺ expression plasmid or vector control were grown in selective media supplemented with 2% glucose at 30°C, washed in ddH₂O, suspended in fresh CSM-uracil-lysine/2% galactose, incubated at 30°C for 16 h, then visualized. (D) Cells expressing GFP-Vps27 and LegC7 were grown as in (C), and localization of GFP-Vps27 was determined.