Figure 5.

Increased Sge1 protein abundance correlates with increased tolerance to IILs. (A) A representative Western blot of different alleles of Myc-tagged Sge1 or actin protein from total cell lysates harvested from the indicated BY strains. Chemiluminescence signal for Sge1-Myc was normalized to actin signal from the same sample. (B) Average normalized Sge1-Myc signals + SEM were plotted from five independent biological replicates. Statistical significance was determined by paired Student’s tests. * P < 0.05. (C) sge1Δ mutant cells containing a plasmid with SGE1-MYC alleles driven by a tetracycline-inducible promoter were cultured in YPD (pH 5) medium containing 0–625 ng/ml doxycycline and 125 mM [C₂C₁im]Cl for 24 hr. Total cell growth was recorded and cells were harvested for total cellular protein lysates after 24 hr. Sge1-Myc protein was quantified with anti-Myc antibodies and normalized for protein loading (see Figure S7 and Materials and Methods). Normalized Sge1-Myc signal from each strain condition was plotted against the total cell growth relative to sge1Δ cells transformed with P_Tet-On-Empty plasmid and grown in 0 ng/ml doxycycline and 125 mM [C₂C₁im]Cl.