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. Author manuscript; available in PMC: 2012 Jul 1.

Published in final edited form as: Mol Microbiol. 2011 Jun 28;81(2):515–527. doi: 10.1111/j.1365-2958.2011.07714.x

Fig. 5 — SIT4 deletion suppresses hydrogen peroxide sensitivity of isc1Δ cells. S. cerevisiae BY4741, isc1Δ, sit4Δ and sit4Δisc1Δ mutant cells were grown in YPD medium to the exponential phase (OD₆₀₀=0.6) (white bars) and exposed to 1.5 mM H₂O₂ for 30 min (black bars). (A) Cell viability was determined by standard dilution plate counts and expressed as the percentage of the colony-forming units of non-stressed cells. (B) Intracellular oxidation. Cells were labeled with the molecular probe H₂DCFDA and lysed as described in Materials and Methods. Data was normalized for protein content. (C) Protein carbonylation. Proteins were derivatized with DNPH and slot-blotted into a PVDF membrane. Immunodetection was performed using an anti-DNP antibody, as described in Materials and Methods. Quantitative analysis of total protein carbonyl content was performed by densitometry using data taken from the same membrane. Values are means ± SD of three independent experiments. *p<0.05, **p<0.01.

Fig. 5 — SIT4 deletion suppresses hydrogen peroxide sensitivity of isc1Δ cells. S. cerevisiae BY4741, isc1Δ, sit4Δ and sit4Δisc1Δ mutant cells were grown in YPD medium to the exponential phase (OD₆₀₀=0.6) (white bars) and exposed to 1.5 mM H₂O₂ for 30 min (black bars). (A) Cell viability was determined by standard dilution plate counts and expressed as the percentage of the colony-forming units of non-stressed cells. (B) Intracellular oxidation. Cells were labeled with the molecular probe H₂DCFDA and lysed as described in Materials and Methods. Data was normalized for protein content. (C) Protein carbonylation. Proteins were derivatized with DNPH and slot-blotted into a PVDF membrane. Immunodetection was performed using an anti-DNP antibody, as described in Materials and Methods. Quantitative analysis of total protein carbonyl content was performed by densitometry using data taken from the same membrane. Values are means ± SD of three independent experiments. *p<0.05, **p<0.01.