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. 2020 Aug 13;10:13740. doi: 10.1038/s41598-020-70793-6

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© The Author(s) 2020

Open AccessThis 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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Functionality of bicistronic lentiviral constructs in vitro. (A) T1934.4 cells were transduced using MOI 50 with the dLck-lentivirus or the CD3δ-lentivirus. eGFP and mCherry expression were analyzed 7 days post lentiviral transduction. Transduced cells (right and left) are double positive for eGFP and mCherry compared to non-transduced cells (middle). (B) Primary murine spleen T-cells were infected with the respective lentivirus as depicted and expanded in vitro using CD3xCD28 costimulation for 3 days in a single transduction experiments. The dot plots depict eGFP and mCherry expression for dLck-lentivirus (middle) and CD3δ-virus (right) treated T-cells gated on mCherry positive and negative subpopulations.