Figure 5.

Overlapping requirements for Ski7 and Dom34/Hbs1 during oxidative stress conditions. (A) Sensitivity to oxidative stress was determined by spotting strains onto minimal media containing various concentrations of H₂O₂. Results are shown for the wild-type, ski7, dom34, hbs1, ski7 dom34 and ski7 hbs1 mutant strains following 3 days growth on 2.5 mM H₂O₂. (B) Viability analysis is shown for the same strains grown to exponential phase in minimal media and treated with 2 mM H₂O₂ for 1 h. Cells were diluted and plated in triplicate onto YEPD medium to monitor cell viability. Percent survival is expressed relative to the untreated control cultures. (C) Protein was isolated from the wild-type and indicated mutant strains expressing Protein A-non-stop protein and analyzed by western blotting. Blots were probed with a Protein A antibody (NSD) and an actin antibody as a loading control. Quantification is shown for Protein A concentrations relative to actin. Data are the means of three independent biological repeats and error bars denote SD. (D) Quantitative RT-PCR of Protein A-non-stop mRNA levels. Quantification is shown for Protein A mRNA concentrations relative to actin mRNA. Data are the means of three independent biological repeats and error bars denote SD.