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. 2019 Apr 23;9:6467. doi: 10.1038/s41598-019-42914-3

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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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Induction of eGFP expression from a CEM-SS cell line containing a stably integrated single copy of the reporter plasmid. CEM-SS cells were transduced with an eGFP-mCherry HIV-derived construct and a clonal cell line was created by limiting dilution. Thus, almost all cells show a uniformity of mCherry expression (Plot a-left). The cell line was subsequently transduced by a pMSCV-Rev-IRES-TagBFP and fluorescence was measured for all three markers. Untransduced cells are shown on the left, cells transduced with the MSCV-derived Rev/BFP expressing vector are shown on the right. Plot a (right) shows that transduction caused about 8.27% of the cells to express TagBFP. Plot b (right) shows that transduction caused about 8.79% of the cells to express eGFP. Plot c (right) shows that there is a linear relationship between the BFP and eGFP signal, indicating a strong correlation between Rev levels in a cell (BFP) and expression of the unspliced mRNA isoform (eGFP).