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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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Schematic overview of lentiviral constructs and experimental study design. (A) A bicistronic lentivirus: the global mPGK-promoter drives mCherry expression in all transduced cells, whereas the dLck- or CD3δ-promoter regulates eGFP-expression restricted to T-cells (cell-specific-promoter). (B) Experimental design: CD105⁺/Sca-1⁺ enriched BM cells were isolated from donor mice, transduced with the lentiviral constructs at day 1 and 2, followed by an enrichment of mCherry⁺ cells by FACS-sorting on day 7. Irradiated recipient mice were transplanted with mCherry⁺ HSCs. 24 weeks after HSC-transplantation. Thioglycolate mediated peritonitis was induced and samples were harvested for flow cytometric analysis 3 days later.