Figure 5. The role of ATM and H2AX in RMD.

• A, B
  SSA/RMD and BIR/SSA are both dependent on ATM. SSA/RMD and BIR/SSA frequencies in U2OS (EGFP‐SSA‐3427‐17) cells (A) and BIR/RMD in mES cells carrying the RMD‐GFP reporter (B) were determined after the addition of ATM inhibitor (20 mM ku55933) or DMSO to the cell culture medium 24 h before transfection of the indicated sgRNA/Cas9 (A and B), or in ATM KO mES cells carrying the RMD‐GFP reporter (right).
• C, D
  Dependence of RMD on H2AX. SSA/RMD and BIR/RMD frequencies in U2OS (EGFP‐SSA‐3427‐17) cells (C) and BIR/RMD in mES cells carrying the RMD‐GFP reporter (D) were determined using the indicated sgRNA/Cas9 after depletion of H2AX by lentivirus‐expressed shRNA (C), or by transfection with H2AX siRNA or in RAD52 KO cells (D). Depletion of H2AX by shRNA in (C) and by siRNA in (D) was determined by qPCR shown in Appendix Fig S8B and C, respectively.

Data information: Error bars represent standard deviation (SD) of at least three independent experiments. *P < 0.05, **P < 0.01, ns, not significant, two‐tailed non‐paired t‐test.