Fig. 1 | Scientific Reports

Fig. 1

From: High-throughput DNA repair monitoring in Saccharomyces cerevisiae suggests SSB- and DSB-induced chromatin reconfiguration

Fig. 1

Real-time tracking and quantification of MATa-stk switching to MATα. (A) Mating-type switching MATa-stk strain, stably expressing inducible HO-endonuclease (by galactose), and the switching fluorescent reporter CFP. Induction of HO-endonuclease results in a mixed population with either intact DNA or damaged DNA (SSB or DSB). Cells that repaired their DNA by switching, express CFP in the nucleus. (B) MATa-stk locus quantification by qPCR; disappearance of this locus quantifies the DNA damage, while its reappearance corresponds to non-switching repair. Tracking and quantifying switching by (C) qPCR of MATα locus, and CFP as fluorescent marker in (D) IFC, and (E) FC. The ~ 90 min difference between switching detection by qPCR (C) vs. flow methods (D), (E), is consistent with CFP maturation time of ~ 90 min (see table S1). All experiments obtained significant switching levels (p-value < < 0.01) by comparing induced to uninduced (0% galactose) cultures using either N-way or two-way ANOVA.

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