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. 2016 Sep 28;1(5):e00250-16. doi: 10.1128/mSphere.00250-16

FIG 4  .

FIG 4 

Rom2 protein levels in the ccr4Δ and khd1Δ ccr4Δ mutant strains. (A, top) Schematic representation of the regulatory elements in the ROM2myc construct. (Bottom) Rom2myc protein levels in wild-type, khd1Δ ccr4Δ, khd1Δ, and ccr4Δ cells. Wild-type (180-3B-4A), khd1Δ ccr4Δ (180-3B-1A), ccr4Δ (180-3B-7C), and khd1Δ (180-3B-7D) cells harboring the ROM2myc construct were cultured to mid-logarithmic phase in YPD medium and collected, and total protein was prepared. The Rom2myc proteins were quantified by Western blotting as described in Materials and Methods. Mcm2 protein was included as a quantity control. The protein levels shown below the lanes are indicated as percentages of the wild-type levels and represent the means ± standard deviations from three independent experiments. (B, top) Schematic representation of the regulatory elements in the pGAL-HA-ROM2 construct. (Bottom) HA-ROM2 mRNA levels in wild-type, khd1Δ ccr4Δ, khd1Δ, and ccr4Δ cells. Wild-type (c1H-1A), khd1Δ ccr4Δ (c1H-1B), khd1Δ (c1H-1C), and ccr4Δ (c1H-1D) cells harboring pGAL-HA-ROM2 plasmid were cultured to mid-logarithmic phase in SG−Ura medium and collected, and total RNA was prepared. The HA-ROM2 transcripts were quantified by Northern blotting as described in Materials and Methods. ACT1 mRNA was included as a quantity control. The mRNA levels are indicated as percentages of wild-type levels and represent the means ± standard deviations from three independent experiments. (C) HA-Rom2 protein levels in wild-type, khd1Δ ccr4Δ, khd1Δ, and ccr4Δ cells. Wild-type (c1H-1A), khd1Δ ccr4Δ (c1H-1B), khd1Δ (c1H-1C), and ccr4Δ (c1H-1D) cells harboring pGAL-HA-ROM2 plasmid were cultured to mid-logarithmic phase in SG−Ura medium and collected, and total protein was prepared. The HA-Rom2 proteins were quantified by Western blotting as described in Materials and Methods. Mcm2 protein was included as a quantity control. The protein levels are indicated as percentages of wild-type levels and represent the means ± standard deviations from three independent experiments.