Fig. 1. Role of Ncr1p in hydrogen peroxide resistance.
S. cerevisiae BY4741 and ncr1Δ::KanMX4 cells were grown in SC-glucose medium to exponential phase (O.D.600nm=0.6) exposed to 1.5 mM H2O2 for 1 hour.
A. Cellular viability was measured as the percentage of the colony-forming unit (treated cells vs non-stressed cells). Values are mean ± SD of at least three independent experiments. ***p<0.001, unpaired Student’s t-test.
B. Protein carbonylation. Proteins were derivatized with DNPH and slot-blotted into a PVDF membrane. Immunodetection was performed using an anti-DNP antibody. Quantitative analysis of total protein carbonyl content was performed by densitometry using data taken from the same membrane. Values are mean ± SD of at least three independent experiments.****p<0.0001, **p<0.01; Two-way ANOVA and Bonferroni test.
C. Lipid peroxidation. Cellular extracts were prepared and TBARS quantification was performed as described in Experimental procedures. Values are mean ± SD of at least three independent experiments. ***p<0.001; Two-way ANOVA and Bonferroni test.
D. Intracellular levels of superoxide radicals were analyzed by flow cytometry, using DHE as probe. Values are mean ± SD of at least three independent experiments. ****p<0.0001, **p<0.01; Two-way ANOVA and Bonferroni test.