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We also overexpressed Artemis in control cells and observed that D37N-mutant Artemis, but not WT Artemis, increased the γH2AX foci level and decreased the Rad51 foci level.
Furthermore, expression of WT Artemis, but not D37N Artemis, restored the Rad51 foci level of Artemis-deficient fibroblasts.
Round spermatids showed highly increased γH2AX foci level at 5, 24, and 48 h after IR in comparison with normal somatic cells in lung, kidney, and brain.
Given the heightened p-H2AX foci level and sustained PARP-1 activation state observed in cyclin D1b-expressing lines, these cells appear reliant on PARP-1 signaling cascades for genomic maintenance.
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Moreover, depletion of Ku80 or Lig4 does not substantially affect BrdU foci levels.
In contrast, KU55933 increased foci levels in Lig4−/− MEFs in G2.
Indeed, pre-treatment with α-LA at concentrations >1 μM reduced γ-H2AX foci levels to WT levels.
Further, KU55933 did not affect foci levels in Rad54−/− MEFs in G2, demonstrating epistasis between ATM and Rad54 in G2.
(F ) Rescue of control DSB foci levels after knockdown of FMN2 by overexpression of hairpin-resistant mutant of mCherry-FMN2.
Interestingly, α-LA treatment had no effect on γ-H2AX foci levels in WT-LCLs at the concentrations tested.
The pre- and post-CT exposure foci levels are presented in Fig. 1 for every individual patient.
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