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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 PFT-μ and ZVAD-fmk. (a) HPBLs were treated with 20 μM PFT-μ for 5 h or 100 μM ZVAD-fmk for 1 h before X-irradiation (2 Gy). The levels of p-p53 expression (left upper panel) and cleaved caspase-3 expression (left bottom panel) in HPBLs at indicated time points post irradiation were examined by western blotting. Furthermore, pan-nuclear γH2AX and TUNEL-positive cells were counted at 24 h post irradiation. (b) HPBLs was treated with 20 μM PFT-μ for 5 h before X-irradiation. Pan-nuclear p-ATM- and p-DNA-PKcs-positive cells were counted at 6 h post irradiation (upper panel). γH2AX, p-ATM and p-DNA-PKcs foci were measured in HPBLs at 0.5 h (left bottom panel) and 4 h (right bottom panel) post irradiation. A total of 1000 HPBLs from each sample were examined for quantitation of the p-ATM, p-DNA-PKcs and γH2AX foci, and 2000–5000 HPBLs from each sample were examined to calculate the percentage of pan-nuclear γH2AX, p-ATM, p-DNA-PKcs and TUNEL-positive cells. The values shown in panel (a) and panel (b) 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 with/without inhibitor treatment; ^## P<0.01 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.