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. 2019 Sep 3;9:12680. doi: 10.1038/s41598-019-49061-9

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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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HeLa cells display an increased cellular proliferation and viability when p53 DBD Nbs are expressed. An XTT assay was performed to evaluate whether or not the p53 DBD Nb-induced stabilisation of p53 levels affects the cell viability of HeLa cells. The cells were transfected with FLAG-tagged p53 DBD Nbs or a FLAG-tagged GFP Nb (control). Thereafter, the transfected cells received (or did not receive) an additional treatment with 10 µM SAHA or 1 µM staurosporine for a duration of 20 h. The graph depicts the mean net absorbance of the formazan dye measured at 475 nm (±SEM) of triplicate samples in 4 independent experiments. Statistical analysis was performed by means of a two-way ANOVA with a Bonferroni post-test. A significant higher absorbance was measured for HeLa cells expressing the p53 DBD Nbs (p < 0.001). This result implicates that expression of p53 DBD Nbs coincided with an increased proliferation and viability of HeLa cells. When the cells were co-treated with 10 µM SAHA, there was a decline in cellular proliferation. However, there was still a significant difference between HeLa cells expressing a p53 DBD Nb and the control condition (p < 0.001). The cell viability was dramatically reduced for all conditions when the cells were co-treated with 1 µM staurosporine.