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. 2016 Mar 21;2:16011. doi: 10.1038/cddiscovery.2016.11

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Copyright © 2016 Cell Death Differentiation Association

This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/

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Inhibition of X-irradiation-induced pan-nuclear γH2AX response and apoptosis in resting HPBLs by KU55933. (a) HPBLs were treated with 10 μM KU55933 for 2 h before X-irradiation (0.5 or 5 Gy). γH2AX, p-ATM and p-DNA-PKcs foci were measured in 500–1000 HPBLs from each sample. (b) HPBLs were treated with 10 μM KU55933 for 2 h before X-irradiation (5 Gy). Pan-nuclear p-ATM and p-DNA-PKcs cells (left panel) were counted, and western blotting assays (right panel) were performed to examine the expression of p-ATM/ATM, p-DNA-PKcs/DNA-PKcs and p-p53/p53 proteins at the indicated time points after irradiation. (c) HPBLs were treated with 10 μM KU55933 for 2 h before X-irradiation (0.5 or 5 Gy). Pan-nuclear γH2AX and TUNEL-positive cells were measured, and their representative images were captured at 24 h after X-irradiation; γH2AX was labeled in red and TUNEL in green, and nuclei were stained in blue with DAPI. A total of 2000–5000 HPBLs from each sample were examined to determine the percentage of the pan-nuclear γH2AX, p-ATM, p-DNA-PKcs and TUNEL-positive cells. ×100 magnification, ×1.0 zoom. The values shown in panel (a), panel (b) and panel (c) represent the means±S.D. obtained from three to four donors. *P<0.05, **P<0.01 and ***P<0.001 compared with corresponding non-irradiated HPBLs; ^## P<0.01 and ^### P<0.001 compared with corresponding HPBLs exposed to 0.5 Gy X-ray irradiation with/without inhibitor treatment; ^§ P<0.05, ^§§ P<0.01 and ^§§§ P<0.01 compared with corresponding HPBLs without inhibitor treatment.