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. 2021 Apr 12;40(10):e104847. doi: 10.15252/embj.2020104847

Figure 2. Rad52 and Rad59 suppress BIR during MAT switching.

Figure 2

  1. Schematic of the H‐1400 assay. A DSB is induced at MATa and repaired by recombination with HMLα‐inc. There is ~ 1,400 bp homology between MATa and HMLα‐inc sequences on the second end (green box).
  2. Representative Southern blots showing DSB repair products in WT, rad59Δ, rad52‐R70A, and rad59Δ rad52‐R70A cells. DNA was digested with XhoI and EcoRI and probed with a Z sequence (yellow box). A detailed map of restriction cut sites and expected size for different repair products are illustrated in Appendix Fig S1.
  3. Percentage of BIR among all repair products by 6 h (mean ± SD; n = 3).
  4. Viability of indicated strains (mean ± SD; n = 3).
  5. Graphs show analysis of repair efficiency compared to uncut parental band (left) (mean ± SD; n = 3) and repair efficiency compared to WT (right) at 6 h after DSB induction (right). See Fig 1F for details.
  6. Schematic of the H‐2100 no‐gap assay. A DSB is induced at MATα and repaired by recombination with HMLα‐inc. There is ~ 2,100 bp homology between MATα and HMLα‐inc sequences on the second end (green box).
  7. Representative Southern blots showing DSB repair products in WT, rad59Δ, and rad52‐R70A. DNA was digested with XhoI and EcoRI and probed with a Z sequence (yellow box). A detailed map of restriction cut site and expected size for different repair products are illustrated in Appendix Fig S1.
  8. Percentage of BIR product among repair products by 6 h are shown (mean ± SD; n = 3).
  9. Viability of indicated strains (mean ± SD; n = 3).
  10. Graphs show analysis of repair efficiency compared to uncut parental band (left) (mean ± SD; n = 3) and repair efficiency compared to WT (right) at 6 h after DSB induction (right). See Fig 1F for details.

Data information: Welch’s unpaired t‐test was used to determine the P‐value in all panels. *P‐value 0.01 to 0.05, significant; **P‐value 0.001 to 0.01, very significant; ***P‐value 0.0001 to 0.001, extremely significant; ****P < 0.0001, extremely significant; P ≥ 0.05, not significant (ns).