Yeast cells were first treated under the same conditions as mentioned above. Yeast cells were irradiated with a UV dose of 106.5 J/m2 UV-C (254 nm) under a Vilber VL-6.C filtered lamp (Thermo Fisher Scientific, Villebon-sur-Yvette, France), and incubated at 28 °C with orbital shaking at 120 rpm in the dark in complete 2.0% (w/v) glucose YPD medium (Sigma Aldrich, Saint-Quentin Fallavier, France) as previously described [65 (link)]. The same conditions were used to grow non-irradiated cells. Hour 0 of the oxidative stress experiment was considered irradiation.
The dihydrorhodamine-123 (DHR-123) fluorescent dye (Sigma-Aldrich, Saint-Quentin Fallavier, France) was used to assess the quantity of reactive oxygen and nitrogen species. Approximately 108 yeast cells were washed twice in PBS, resuspended in PBS containing 0.4 M DHR-123, and incubated for 10 min in the dark at 28 °C in the presence of extract, RES, or DMSO (control cells). The fluorescence signal (ex = 505 nm, em = 535 nm) was measured using the VersaFluor Fluorimeter after two washes with PBS (Biorad, Marnes-la-Coquette, France).
The dihydrorhodamine-123 (DHR-123) fluorescent dye (Sigma-Aldrich, Saint-Quentin Fallavier, France) was used to assess the quantity of reactive oxygen and nitrogen species. Approximately 108 yeast cells were washed twice in PBS, resuspended in PBS containing 0.4 M DHR-123, and incubated for 10 min in the dark at 28 °C in the presence of extract, RES, or DMSO (control cells). The fluorescence signal (ex = 505 nm, em = 535 nm) was measured using the VersaFluor Fluorimeter after two washes with PBS (Biorad, Marnes-la-Coquette, France).
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