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. 2021 Nov 24;41(1):e108813. doi: 10.15252/embj.2021108813

Figure EV1. Sir3 overexpression limits resection and increases error‐prone NHEJ.

Figure EV1

  1. Survival frequencies and characterization of the repair events after induction of a DSB at LYS2 in absence of recombination substrate.
  2. Survival frequencies and characterization of the repair events after induction of a DSB at TEL6R in absence of recombination substrate.
  3. DNA levels measured at 1 kb from the I‐SceI cut site at LYS2 after 2 h DSB induction by qPCR in WT and sae2Δ cells expressing or not high levels of Sir3p (oeSir3 and WT, respectively). DNA levels were normalized to DNA levels at the OGG1 locus and corrected for differences in DSB cleavage efficiency (see Materials and Methods for details). Error bars represent the standard deviation (SD) of three independent experiments.

Data information: (A, B) NHEJ stands for error‐prone end‐joining events detected by a PCR product that cannot be cleaved in vitro by I‐SceI. No DSB corresponds to survivors giving a PCR product that can be cleaved by I‐SceI in vitro showing that they failed to induce I‐SceI. Other gathers survivors in which no PCR product was obtained suggesting that repair occurred through other mechanisms. PCR products corresponding to NHEJ events were sequenced and exhibit patterns typical of NHEJ repair (rejoining with 1‐ to 9‐bp deletion between sequences showing no or limited homology).