Glutamax 1 25 mm hepes

Yeast cells of selected strains were cultivated in RPMI 2% glucose, pelleted at 3,220 × g for 10 min at 4°C, washed twice in PBS, and resuspended at a concentration of 1.25 × 10⁷ /mL in fresh RPMI 1640 GlutaMAX-I 25 mM HEPES (Gibco) containing 0.4 mM d-luciferin without addition of an oxidative stressor (control) or supplemented with 20 mM hydrogen peroxide (H₂O₂; Sigma) or 8 mM tert-butylhydroperoxide (tBOOH; Sigma). The yeasts were incubated at 37°C and 5% CO₂. The relative light units per second (RLU/s) were measured after 8 h using a TECAN Spark plate reader set at 37°C and 5% CO₂ in the air space. The assays were performed at least two times in triplicates. The mean of luminescence and standard deviations were plotted. A statistically significant difference between Δku80 and Δku80_Δpks1 was calculated using one-way ANOVA in GraphPad Prism version 9 for Windows.

Yeast cells of selected strains were cultivated in RPMI 2% glucose, pelleted at 3,220 × g for 10 min at 4°C, washed twice in PBS, and resuspended at a concentration of 1.25 × 10⁷ /mL in fresh RPMI 1640 GlutaMAX-I 25 mM HEPES (Gibco) containing 0.4 mM d-luciferin without addition of an oxidative stressor (control) or supplemented with 20 mM hydrogen peroxide (H₂O₂; Sigma) or 8 mM tert-butylhydroperoxide (tBOOH; Sigma). The yeasts were incubated at 37°C and 5% CO₂. The relative light units per second (RLU/s) were measured after 8 h using a TECAN Spark plate reader set at 37°C and 5% CO₂ in the air space. The assays were performed at least two times in triplicates. The mean of luminescence and standard deviations were plotted. A statistically significant difference between Δku80 and Δku80_Δpks1 was calculated using one-way ANOVA in GraphPad Prism version 9 for Windows.