Expression levels of plasmid expressed Svf1 are slightly altered. (a) Expression levels of Svf1-GFP constructs used in this study. Equal amounts of cells were lysed and analyzed by Western blotting using antibodies against the GFP-tag or Pgk1 as a loading control. NC = negative control/ WT strain; end. WT = Svf1 GFP tagged endogenously with GFP; pl. WT = Svf1-GFP expressed under its endogenous promoter from an integrative plasmid; V12D = Svf1_V12D-GFP expressed under its endogenous promoter from an integrative plasmid; G7A/G8A = Svf1_G7A/G8A-GFP expressed under its endogenous promoter from an integrative plasmid; L2E = Svf1_L2E-GFP expressed under its endogenous promoter from an integrative plasmid. (b) Helical wheel representation of the first 18 amino acids with the hydrophobic amino acids shown in yellow and the hydrophobic moment shown by the arrow and expressed in µH above the arrow for the L2E mutant with the exchange of the hydrophobic leucine to the charged glutamate (red). Source data are available for this figure: SourceData FS3.