CK2 depletion results in decreased DNA-PKcs phosphorylation in M059K cells following induction of DNA double-strand break. A. Cells were treated as described in Figure 1. Whole cell extracts were employed for Western blot analysis of the indicated proteins. Detection of β-actin was used as loading control. B. Cells were treated essentially as described in Figure 1 except that DNA damage was induced by cell exposure to 10 Gy ionizing radiation. After 1 hour, cells were harvested and total cell lysates were prepared for Western blot analysis. C. M059K cells were treated with 33 μM cisplatin (cisPt) or exposed to 20 J/m2 UV irradiation for the indicated time-points. Cell extract from control and treated cells was prepared and analyzed by Western blot employing antibodies against DNA-PKcs or its phosphorylated forms as indicated in the figure. D. Experiments were performed as described above and in Figure 1 except that cells were treated with cisPt (33 μM, 8 hours incubation) or exposed to UV (20 J/m2, 8 hours recovery) as indicated in the figure. Whole cell lysates were prepared for the analysis by Western blot of DNA-PKcs phosphorylation at S2056, T2609 and T2647. Experiments were performed at least three times obtaining similar results. Data from one representative experiment are shown.
Olsen et al. BMC Molecular Biology 2012 13:7 doi:10.1186/1471-2199-13-7