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. 2020 Mar 10;21(5):1894. doi: 10.3390/ijms21051894

Figure 3.

Figure 3

Changes in the expression levels of different alleles of Med15 treated with MCHM. (A) Protein levels of Med15-13xMyc expressed from a plasmid in BY4741 med15 yeast. The yeast was grown in selective media until mid-log and then shifted to 550 ppm MCHM for 90 minutes. Med15-13xMyc was immunoprecipitated from equal amounts of protein extract. (B) mRNA levels of MED15 expressed from a plasmid in BY4741 med15 yeast normalized to ACT1 mRNA. Transcript levels were extracted from RNAseq data. The yeast was grown in YPD (with G418) or YM (yeast minimal media supplemented with histidine, leucine and methionine (HLM) and then treated with 550 ppm MCHM for 30 minutes. (C) Western blot of Med15-Myc immunoprecipitated from BY4741 carrying YJM789 and S288c alleles of Med15 on single copy plasmids from yeast grown in YPD at 0, 15, 30, and 90 minutes after the addition of cycloheximide. The total lysate was run separately and Pgk1 was blotted as a loading control.