DNA-PKcs−/−, Ku70−/−, and wild-type mice on 129 genetic backgrounds were bred as previously reported [46 (link), 47 (link)]. Mouse embryonic fibroblast (MEF) cells from DNA-PKcs−/−, Ku70−/−, and wild-type animals were isolated from 13.5-day-old mouse embryos, respectively. The genotype of the embryos was determined by PCR, which distinguished the endogenous from the targeted DNA-PKcs allele or Ku70 allele.
NU7441 (Tocris Bioscience), a DNA-PK inhibitor was dissolved in dimethylsulfoxide (DMSO) for 5 mmol/L stocks and stored at −20°C. All drugs were added to cells to a final DMSO concentration of 0.5%. Cells were exposed to X-rays generated by a RAD SOURCE RS2000 irradiator (Rad Source Technologies, Inc.) operating at 25 mA, with a 0.3-mm Al filter and effective photon energy of 160 kV. The dose rate at an irradiation distance of 48.6 cm was 1.31 Gy/min.
NU7441 (Tocris Bioscience), a DNA-PK inhibitor was dissolved in dimethylsulfoxide (DMSO) for 5 mmol/L stocks and stored at −20°C. All drugs were added to cells to a final DMSO concentration of 0.5%. Cells were exposed to X-rays generated by a RAD SOURCE RS2000 irradiator (Rad Source Technologies, Inc.) operating at 25 mA, with a 0.3-mm Al filter and effective photon energy of 160 kV. The dose rate at an irradiation distance of 48.6 cm was 1.31 Gy/min.
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