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. 2003 Aug;23(16):5836–5848. doi: 10.1128/MCB.23.16.5836-5848.2003

FIG. 4.

FIG. 4.

An ABCDE mutant with aspartic acid substitutions is partially competent in NHEJ. (A) Immunoblot analyses of whole-cell extracts from V3 transfectants expressing wild-type (wt) DNA-PKcs (lane 1), vector alone (lane 2), mutant ABCDE (lane 3), the ST→D mutant (clone 5), (lane 4), and the ST→D mutant (clone 11) (lane 5). (B) Whole-cell extracts (500 μg) prepared from V3 cells transfected with either vector alone, wt DNA-PKcs, mutant ABCDE, or one of two independent clones (clones 5 and 11) expressing the ST→D mutation were assayed for DNA-PK activity. Phosphorylation of the p53 substrate was assessed after 60 min. Each cell extract was tested in duplicate, and three independent extracts were tested for each cell line. Error bars, standard deviations. (C) Radiation resistance of V3 transfectants expressing wt DNA-PKcs, vector alone, mutant ABCDE, or the ST→D mutant. Data are presented as percent survival relative to that of nonirradiated controls (set at 100%). Error bars, standard errors of the means of three separate experiments.