Table 1.

Relevant parameters for ATR activation upon a subset of genotoxic treatments

ParameterApproachFigureNTMMC (200 nM)APH (100 nM)CPT (25 nM)HU (0.5 mM)
Fork reversalEM analysis3, B and C−/+++++++++
Fork slowingDNA fiber spreading1 A++++++++
Impaired DNA synthesisEdU incorporation (FACS)4 D−/+++++++
ssDNA at forksEM analysis2, A and B; S2; and S3++++
Total exposed ssDNANative BrdU staining4 B++++
ATR signaling at forksiPOND γ-H2AX6 B−/+ND+++++
ATR signaling totalWB pCHK14 A++++
ATR signaling totalIF/FACS γ-H2AX4, B–D++++
  • Parameters were assessed by different investigation methods, as displayed in the indicated figures. −/+, +, ++, and +++ indicate increasingly clear phenotypes. IF, immunofluorescence; NT, not treated; WB, Western blot.