Fig2

Fig 2. Analyses of cell rescue proteins, redox state, and protein oxidation under oxidative conditions. (A) Expression changes in antioxidant and metabolic enzymes in mid-log phase yeast cells exposed to 20 mM H2O2 for 1 h with shaking. Tubulin (Tub) was used as a loading control. (B) Hydroperoxide levels in TC cells in the absence (red bar) and presence (green bar) of 20 mM H2O2 were assessed using FOX reagent and were calculated relative to that in WT cells grown under normal conditions, which was set to 100%. (C) Mid-log phase yeast were exposed to 20 mM H2O2 for 1 h at 28°C with shaking. Redox state was analyzed by measuring DCFHDA oxidation as an indicator of cytosolic ROS. (D) Sensitivity of mutants (sod1Δ, tsa1Δ, por1Δ, and por2Δ) to oxidative stress. Yeast cells (OD600 ≈ 1.0) were exposed to 10 mM H2O2 for 1 h at 28°C with shaking, serially diluted with YPD medium, spotted onto YPD agar plates, and incubated for 2–3 days. (E) Expression changes in molecular chaperones in mid-log phase yeast cells exposed to 20 mM H2O2 for 1 h with shaking. Tubulin (Tub) was used as a loading control. (F) Protein carbonylation in yeast cells exposed to 20 mM H2O2 for 1 h was calculated relative to that in WT cells under normal conditions, which was set to 100%. Red bar, normal conditions; green bar, H2O2 treatment; WT, yeast cells with an empty vector; TC, OsMDHAR-expressing yeast cells; N, normal conditions; S, H2O2 treatment.