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. 2020 Mar 20;11:1509. doi: 10.1038/s41467-020-15140-z

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© The Author(s) 2020

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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Fig. 2 — a DAP restores expression of the endogenous TP53 gene if it carries a UGA nonsense mutation but not if it carries a UAA or UAG nonsense mutation. Calu-6 (top), Caov-3 (middle), and Caco-2 (bottom) cells were incubated with increasing amounts of DAP or G418 before protein purification and monitoring of the p53 protein level by western blotting. The three leftmost lanes show twofold serial dilutions of untreated HeLa cell extract. The molecular weight is indicated to the left of each gel. b Schematic representation of the construct used to measure the function of p53. The construct carries a cDNA encoding the firefly luciferase under a promoter having two p53-responsive elements (underlined). The arrow indicates the transcription start. c Luciferase activity in Calu-6 (upper panel) and HeLa cells (lower panel) transfected with the construct described in b and treated with DMSO, DAP, or G418. The luciferase activity depends on the amount of functional p53 protein. d Level of p21 mRNA measured by quantitative RT-PCR in Calu-6 (upper panel) and HeLa cells (lower panel) treated with DMSO, DAP, or G418. The level of p21 mRNA is related to the amount of functional p53 protein synthesized in the cell. The results presented in panels b and c are representative of at least two independent experiments., p-values (p) were calculated using Student’s t-test. Source data are provided as a Source Data file.