Measuring ROS Production in Yeast Cells

The production of reactive oxygen species (ROS) was assessed using dihydrorhodamine 123 (DHR; Sigma-Aldrich) as described previously^{60 (link)}. Cells were grown in YP galactose medium until mid exponential phase, a sample was collected and 100 mM glucose was added after which a new sample was taken. 5 µg/mL DHR was added to the cells and incubated for 2 h. ROS production was visualized using fluorescence microscopy (Axioplan 2 imaging, Zeiss).

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Peeters K., Van Leemputte F., Fischer B., Bonini B.M., Quezada H., Tsytlonok M., Haesen D., Vanthienen W., Bernardes N., Gonzalez-Blas C.B., Janssens V., Tompa P., Versées W, & Thevelein J.M. (2017). Fructose-1,6-bisphosphate couples glycolytic flux to activation of Ras. Nature Communications, 8, 922.

Publication 2017

dihydrorhodamine 123 (DHR; Axioplan 2 imaging

Cells Dihydrorhodamine 123 Fluorescence microscopy Galactose Glucose Reactive oxygen species

Corresponding Organization :

Other organizations : KU Leuven, VIB-VUB Center for Structural Biology, VIB-KU Leuven Center for Microbiology

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Variable analysis

independent variables

Addition of 100 mM glucose

dependent variables

Production of reactive oxygen species (ROS)

control variables

Cells grown in YP galactose medium until mid exponential phase
Incubation of cells with 5 µg/mL dihydrorhodamine 123 (DHR) for 2 h

controls

Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned

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