Fig. 3: Transcription of the msmeg_1357-56 operon is increased upon genotoxic and oxidative stress. | Communications Biology

Fig. 3: Transcription of the msmeg_1357-56 operon is increased upon genotoxic and oxidative stress.

From: Novel WYL domain-containing transcriptional activator acts in response to genotoxic stress in rapidly growing mycobacteria

Fig. 3

a Msm SMR5 wild-type cells were grown to an OD600 of 0.5 at 37 °C and stressed with 10 mM diamide, 7 mM H2O2, 80 ng/mL MMC, 15 mJ/cm2 UV or 5 µg/mL zeocin for 30 min. RT-qPCR analysis revealed that msmeg_1357 (lavender) and msmeg_1356 (purple) transcript levels are upregulated among all tested genotoxic conditions but most strongly upon H2O2 and zeocin stress. Relative transcript levels were calculated by normalizing CT values of msmeg_1357 and msmeg_1356 against sigA. Accordingly, 2-ΔCT values were calculated and normalized to untreated cells. b Msm SMR5 wild-type cells were cultured as described before and treated with 0.2, 1 or 5 mM H2O2. RT-qPCR analysis shows that msmeg_1357 and msmeg_1356 transcript levels increase in a concentration dependent manner when cells are treated with H2O2. Relative transcript levels were calculated by normalizing CT values of msmeg_1357 and msmeg_1356 against sigA. Next, ΔCT of cells after H2O2 treatment were normalized against ΔCT values of cells before H2O2 treatment. Thus, 2-ΔΔCT values were calculated. Error bars represent the three biological replicates shown as individual data points.

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