Cells were grown at 28°C in SC medium without tryptophan supplemented with 0.6% glucose. Values were normalized to the MSD1 wild-type strain and represented as the mean of at least three values ±SD. (B) Normalized MSD1 protein levels in wild-type and mutant MSD1 strains. Msd1-HA protein levels were determined with Western blot and normalized to Por1 levels. The MSD1:Por1 ratios were normalized to the ratio found in the wild-type strain. Analysis was performed on 3 independent clones for each strain at 28°C. Graphs show average ± SD. Statistical analysis was performed using analysis of variance with Bonferroni correction **p < 0.01, *p < 0.05. Blue bars indicate humanized alleles. MSD1, wild-type; pFL39, empty vector. From left to right in group 1, yLys42Arg/yArg42Gly, yLeu232Val, yPhe487Leu, yAla244Val, yArg157Cys, ySer617Gly, and yIle484Val/yVal484Met correspond to the human mutations Arg58Gly, Leu250Val, Phe484Leu, Ala262Val, Arg179Cys, Ser597Gly, and Val481Met, respectively. From left to right in group 2, yMet608Leu/yLeu608Val and yLys626Met correspond to human mutations Leu588Val and Lys606Met, respectively.