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. 2011 Aug 8;39(21):9224–9237. doi: 10.1093/nar/gkr647

Figure 5.

Figure 5.

IR-induced phosphorylation of PNKP at S114 is transient and ATM dependent while phosphorylation of S126 is transient and requires both DNA-PK and ATM. (A) PNKP knockdown cells (C13) stably expressing shRNA-resistant wt-PNKP were unirradiated (lane 1) or irradiated (10 Gy) and harvested 1, 4, 8, 16 or 24 h post IR. PNKP was immunoprecipitated and immunoblots were probed for phosphorylation at S114, S126 or total PNKP as indicated. (B) Quantitation of immunoblots shown in (A). (C) PNKP knockdown cells (C13) stably expressing shRNA-resistant wt-PNKP were pre-treated with 10 µM KU55933, 10 µM NU7441 or both for 60 min prior to treatment with 10 Gy IR. Cells were harvested after 60 min and PNKP was immunoprecipitated using an antibody to HA, resolved by SDS–PAGE and blotted for phosphoserine 114, phosphoserine 126 or total PNKP as indicated.