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Extra info for Chromosome Aberrations (Reprint of Cytogenetic and Genome Research 2004)
J biol Chem 274:29100–29106 (2000). Brenneman MA, Wagener BM, Miller CA, Allen C, Nickoloff JA: XRCC3 controls the fidelity of homologous recombination: roles for XRCC3 in late stages of recombination. Mol Cell 10:387–395 (2002). Cai Y, Ludeman SM, Wilson LR, Chung AB, Dolan ME: Effect of O6-benzylguanine on nitrogen mustard-induced toxicity, apoptosis, and mutagenicity in Chinese hamster ovary cells. Mol Cancer Ther 1:21–28 (2001). Cole RS: Repair of DNA containing interstrand crosslinks in Escherichia coli: sequential excision and recombination.
The suppressive effect on the MMC-induced SCEs differed among mutants, F 50 % for Rad54–/–, F 80 % for Rad51B–/–, Rad51C–/– and Rad51D–/–, and 90 % or more for Xrcc2–/– and Xrcc3–/– cells. Surprisingly, the frequencies of MMC-induced CAs were also suppressed in Rad54–/–, Rad51B–/– and Rad51C–/– cells. The Rad54–/–/Ku70–/– double knockout mutant responded similarly to the Rad54–/– cells, again indicating no relevance of the NHEJ process in the induction of CAs and SCEs by MMC. It was noteworthy that Rad51D–/–, Xrcc2–/– and Xrcc3–/– were exceptional, in that the 30 Cytogenet Genome Res 104:28–34 (2004) Fig.
Materials and methods Primary neonatal human foreskin fibroblasts were obtained by collagenase digestion and grown to confluence in 150-mm petri dishes in DMEM medium + 10 % fetal calf serum. Medium was replaced two days before irradiation. The cells were washed once in isotonic saline. To prevent desiccation of the cells during irradiation, 2 ml of isotonic saline was added to each plate. Cells were then irradiated with 20 J/m2 of UVC. , Upland, CA, USA). After irradiation, the isotonic salt solution was replaced with fresh medium.