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. 2018 Oct 26;8:15894. doi: 10.1038/s41598-018-34288-9

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© The Author(s) 2018

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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L1 retrotransposition in Rad18-deficient HCT116 cells or Rad18-knockdown 293T cells. (A, B) Rad18^+/+ or Rad18^−/− HCT116 cells (2 × 10⁴ cells/well) were transfected with either pYX014 (A) or pYX017 (B) (100 ng). The luciferase assay was performed three days after transfection in three independent experiments. Graph shows the mean (±SEM) firefly luciferase activity normalized with Renilla luciferase activity. (C) Inhibition of Rad18 protein expression by shRNA-producing lentiviral vector. The results of Western blot analysis of cellular lysates with anti-Rad18 or anti-ß-actin antibody in 293T or HeLa cells expressing shRNA targeted to Rad18 (Rad18KD) as well as in 293T cells transduced with a control lentiviral vector (shCon) are shown. (D) The levels of Rad18 mRNA in Rad18-knockdown 293T or HeLa cells (Rad18KD) or the control cells (shCon) were monitored by real-time LightCycler PCR, respectively. Experiments were done in triplicate, and graph shows the mean percentages (±SEM) of Rad18 mRNA normalized with ß-actin mRNA in the control cells set at 100%. (E,F) Rad18-knockdown 293T (E) or HeLa cells (Rad18 KD) (F) or the control cells (shCon) (2 × 10⁴ cells/well) were transfected with pYX014 (100 ng). Luciferase assays were performed three days after transfection in three independent experiments. Graph shows the mean (±SEM) firefly luciferase activity normalized with Renilla luciferase activity.