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. 2021 Feb 24;11:4480. doi: 10.1038/s41598-021-83979-3

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© The Author(s) 2021

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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/.

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Transient knockdown of TKT slows cell growth and induces cell cycle arrest in H1299 cells irrespective of p53 status. (A) TKT mRNA expression in H1299 parental (p53^null), p53^R273H and p53^R175H cells and JH-EsoAd1 parental (p53^G266E) and p53−/− clone 1 (C1 −/−) following 24 h TKT knockdown with siRNA, normalised to siNTC (siNTC, non-targeting control; siTKT, TKT siRNA). (B) Western blots for TKT in H1299 and JH-EsoAd1 parental cells following TKT knockdown with siRNA for 24, 48, 72 and 96 h, showing actin as a loading control (C) Relative growth of H1299 parental (Par, p53^null), p53^R273H and p53^R175H cells and JH-EsoAd1 parental (p53^G266E), p53−/− clones 1 (C1 −/−) and 2 (C2 −/−) following 96 h TKT knockdown with siRNA. (D) H1299 parental (p53^null) cells + /- N-acetyl-cysteine (NAC, 5 mM) −/−) following 96 h TKT knockdown with siRNA. (C,D) Data are expressed as a percentage of siNTC. (E) Changes in cell cycle distribution in H1299 parental (p53^null) and p53^R273H cells following 96 h TKT knockdown with siRNA. Data represent mean, error bars = SEM, n = 3 for all studies excluding (B,E) where blots and cell cycle analysis are representative of 2 independent experiments. (A,C) Unpaired Student’s t-test (D) One-way ANOVA with Dunnett’s multiple comparison post-test. ns, not significant; *p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001.