<p>A. Mutant <i>MRD1</i> genes were introduced into yeast cells, either as part of a plasmid (P<sub>GAL</sub><i>MRD1</i>+plasmid) or integrated into the genome (P<sub>GAL</sub><i>MRD1</i>+Genomic). In both cases, a WT <i>MRD1</i> gene was present in the genome, controlled by a <i>GAL1</i> promoter. The cells were diluted in steps (10 times for each step) and pipetted onto agar plates (from left to right), containing either galactose or glucose, followed by incubation at 30°C, 37°C or 16°C. Mutants are indicated to the left of each dilution series. The parental strain <i>mrd1-ΔL2</i>::<i>klURA3</i>, (FLY002, See <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0175506#pone.0175506.s003" target="_blank">S1 Table</a>), bearing a non-functional <i>MRD1</i> gene, served as a negative control for cell growth. B. Western blot analyses of the expression of Mrd1p in WT and mutant strains. Extracts from approximately 5x10<sup>5</sup> cells were used. Ponceau staining demonstrated that approximately equal amounts of total proteins was loaded in each well (data not shown). The WT or mutant <i>MRD1</i> genes were either present in the genome or in a plasmid. The protein A part of the HTP tagged proteins was used for detection. WT Mrd1p is 101 kDa and its HTP tag is approximately 17 kDa. Migration of size marker proteins are shown (180, 130, 100, 70, kDa).</p
