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1.
Figure 5 

Figure 5 . From: Regulation of Manganese Antioxidants by Nutrient Sensing Pathways in Saccharomyces cerevisiae.

GIS1 and MSN2/4 have opposing affects on Mn antioxidant protection. The effect of gis1 and msn2/4 mutations on the (A) aerobic lethality of sod1pho80∆ cells, (B) Mn accumulation in sod1pho80∆ cells, and (C and D) Mn-mediated rescue of the sod1∆-linked aerobic lysine auxotrophy in sod1pho80∆ (C) and sod1∆ (D) cells were tested precisely as in Figure 3, A, C, and D, respectively. Details on culture conditions and growth are described in Materials and Methods. All reported values are averages based on triplicate cultures, with the errors representing the standard deviations. Strains employed: sod1∆, AR203; sod1pho80∆, LR156; sod1pho80gis1∆, AR138; sod1pho80msn2/4∆, AR300; sod1gis1∆, AR121; sod1msn2/4∆, AR155.

Amit R. Reddi, et al. Genetics. 2011 December;189(4):1261-1270.
2.
Figure 3 

Figure 3 . From: Regulation of Manganese Antioxidants by Nutrient Sensing Pathways in Saccharomyces cerevisiae.

RIM15 is responsible for the aerobic lethality and poor utility of Mn as an antioxidant in sod1∆ pho80∆ strains. Shown are the effects of a rim15 mutation on (A) aerobic lethality of sod1∆ pho80∆ strains, (B) phosphate accumulation, and (C) Mn accumulation, as described in Figure 2, A, B, and C, respectively. (D) Manganese rescue of the sod1∆-linked aerobic lysine auxotrophy was tested in SC medium lacking lysine and supplemented with the indicated concentrations of MnCl2. Cells were grown as in A except under nonshaking conditions. All reported values are averages based on triplicate cultures, with the errors representing the standard deviations. Strains employed: sod1pho80∆, LR156; sod1pho80rim15∆, AR110.

Amit R. Reddi, et al. Genetics. 2011 December;189(4):1261-1270.
3.
Figure 2 

Figure 2 . From: Regulation of Manganese Antioxidants by Nutrient Sensing Pathways in Saccharomyces cerevisiae.

Activation of the Pho4p transcription factor is not responsible for the aerobic lethality of sod1∆ pho80∆ strains. (A) The effect of a pho4∆ mutation on the aerobic lethality of sod1∆ pho80∆ cells was tested by spotting 104, 103, and 102 cells of the indicated strains onto SCE plates and by growing in air or anaerobically for 3 days. (B) Phosphate content of the indicated strains was measured by molybdate reactivity of the indicated strains grown in SC medium as described in Materials and Methods. (C) Manganese content of the indicated strains was measured by AAS precisely as described in Figure 1C. All reported values are averages based on triplicate cultures, with the errors representing the standard deviations. Strains employed: sod1∆, AR203; sod1pho80∆, LR156; sod1pho80pho4∆, AR106.

Amit R. Reddi, et al. Genetics. 2011 December;189(4):1261-1270.
4.
Figure 4 

Figure 4 . From: Regulation of Manganese Antioxidants by Nutrient Sensing Pathways in Saccharomyces cerevisiae.

Deleting sch9 renders sod1∆ cells oxidatively stressed and unable to utilize Mn for antioxidant protection. (A and B) The effect of sch9∆, tor1∆, and ras2∆ deletions on Mn promotion of aerobic growth was tested in (A) well-aerated SC media and (B) media lacking lysine, as was done as in Figures 1A and 3D, respectively. (C) The effect of sch9∆, sod1∆, and sod1∆ sch9∆ mutations on growth in SCE media in the absence and presence of oxygen and manganese was determined by measuring the o.d.600nm values of shaking cultures after 16 hr of growth, as described in Materials and Methods. All reported values are averages based on triplicate cultures, with the errors representing the standard deviations. Strains employed: sod1∆, AR203; sod1sch9∆, AR163; sch9∆, AR164. sod1tor1∆, AR120; sod1ras2∆, AR158.

Amit R. Reddi, et al. Genetics. 2011 December;189(4):1261-1270.
5.
Figure 7 

Figure 7 . From: Regulation of Manganese Antioxidants by Nutrient Sensing Pathways in Saccharomyces cerevisiae.

GIS1 and MSN2/4 differentially affect the ability of manganese to protect cytosolic but not mitochondrial Fe/S proteins. The indicated strains were grown in SC medium that was supplemented for 6 hr with the designated concentrations of MnCl2 prior to cell lysate preparation and analysis of (A) aconitase and (B) isopropylmalate isomerase (IPMI) activity as described in Materials and Methods. Asterisk denotes a P < 0.05 and reflects the statistical significance between 4 mM Mn treatment and 0 or 2 mM Mn treatment. All reported values are averages based on triplicate cultures, with the errors representing the standard deviations. Strains employed: WT, BY4741; sod1sod2∆, AR142; sod1sod2gis1∆, AR161; sod1sod2msn2/4∆, AR160.

Amit R. Reddi, et al. Genetics. 2011 December;189(4):1261-1270.
6.
Figure 1 

Figure 1 . From: Regulation of Manganese Antioxidants by Nutrient Sensing Pathways in Saccharomyces cerevisiae.

Loss of the Mn–phosphate transporter Pho84p enhances oxygen toxicity in a sod1∆ pho80∆ strain. (A) The indicated sod1∆ strains were seeded in synthetic complete (SC) media at an optical density at 600 nm (o.d.600nm) of 0.05, and grown shaking for 16 hr with the indicated concentrations of MnCl2. Total growth was determined by measuring the o.d.600nm. (B) Manganese toxicity of the indicated strains was determined by growing cells in SC medium supplemented with the indicated concentrations of Mn as described in A under nonshaking conditions. (C) Manganese accumulation of the indicated sod1∆ strains was determined by atomic absorption spectroscopy (AAS) with cells grown in SC medium treated for 6 hr with the indicated concentrations of MnCl2 as described in Materials and Methods. Manganese content is represented as nanomoles per 109 cells. All reported values are averages based on triplicate cultures, with the errors representing the standard deviations. Strains employed: WT, BY4741; pho80∆, LR237; pho80pho84∆, LR154; sod1∆, AR203; sod1pho80∆, LR156; sod1pho80pho84∆, LR178.

Amit R. Reddi, et al. Genetics. 2011 December;189(4):1261-1270.
7.
Figure 6 

Figure 6 . From: Regulation of Manganese Antioxidants by Nutrient Sensing Pathways in Saccharomyces cerevisiae.

GIS1 and MSN2/4 differentially affect manganese-dependent superoxide scavenging activity in SOD-deficient cells. The indicated cells were grown for 6 hr in SC medium that was either (A) not supplemented with metals, or supplemented with (B) 5 mM MnCl2, (C) 5 mM of MnCl2, ZnCl2, CuCl2, or FeCl2, or (D) the indicated concentrations of MnCl2. Whole-cell lysates were prepared and analyzed for total cellular superoxide scavenging activity by the xanthine/xanthine oxidase/XTT assay described in Materials and Methods. One unit is defined as a 50% decrease in the rate of XTT reduction per milligram lysate protein. (A) The asterisk denotes a P < 0.05 and reflects the statistical significance in loss of activity upon a sod2∆ mutation. (B) Where indicated, lysates were incubated for 30 min at room temperature with 1 mM EDTA or at 100° prior to superoxide scavenging activity assays to demonstrate metal chelator sensitivity and heat resistance of SISS activity. Details on culture conditions and the superoxide scavenging assays are described in Materials and Methods. All reported values are averages based on triplicate cultures, with the errors representing the standard deviations. Strains employed: WT, BY4741; sod1∆, AR203; sod1gis1∆, AR121; sod1msn2/4∆, AR155; sod1sod2∆, AR142; sod1sod2gis1∆, AR161; sod1sod2msn2/4∆, AR160.

Amit R. Reddi, et al. Genetics. 2011 December;189(4):1261-1270.

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