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. 2017 Aug 31;7:10213. doi: 10.1038/s41598-017-10222-3

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© The Author(s) 2017

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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The 455UCOE is a core region in the 2.2 kb A2UCOE element preventing the transgene silencing from the SFFV LTR in murine embryonic carcinoma P19 cells. (a) EGFP expression in P19 cells following the viruses transduction. P19 cells were transduced with vectors shown in Fig. 1b, at multiplicity of infection (MOI) of 10. EGFP expression was assessed by flow cytometry at different time points up to 31 days. The numbers above each bar represent vector copy number per cell as determined by Q-PCR. (b) DNA methylation analysis of the SFFV LTR region. Genomic DNA from cells transduced with the SG, the 527UCOE-SG and the 455UCOE-SG vectors was isolated at day 17 post transduction and subjected to methylation analysis by bisulfite conversion and sequencing. The percentages of methylated CpGs to the total of CpGs on the SFFV LTR from randomly selected PCR colonies for each vector are shown. The underlined numbers are the mean of % methylated CpGs from each vector. P values shown are determined by Mann-Whitney U-test (two tailed).