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. 2019 Sep 11;9:13097. doi: 10.1038/s41598-019-49229-3

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© The Author(s) 2019

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Disruption of stressed-induced SMG7 binding to p53 does not significantly affect p53 stabilization, activation or DNA damage response. (a) Western blot analysis of cell extracts from control and irradiated (6 hours) cells of various SMG7 genotypes. (b) α-MDM2 immunoprecipitates from control or irradiated cells were assayed for Ser395 phosphorylation. (c) Cell extracts from control or Doxo-treated (0.4 μM) cells were analyzed by western blot using antibodies as indicated. (d,e) Control or Doxo-treated (0.4 μM, 16 hours) cells were fed with 10 μM CldU for 1 hour and then processed for CldU (green) and DAPI (blue) staining, followed by fluorescence microscopy. Representative images were shown in (d), and quantification of CldU-positive cells from five independent experiments in (e). (*p < 0.05, **p < 0.005; error bars represent SEM). (f) Western blot analysis of cell extracts from control or CPT-treated (1 µM) cells using antibodies as indicated. (g) A working model for SMG7 regulation of p53 stability upon DNA damage.