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. 2018 Nov 2;8:16283. doi: 10.1038/s41598-018-34536-y

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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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Characterisation of HIV-1 Gag(i)mEOS2 wild type and mutants expressed in Jurkat T cells. (a) Schematic representation of HIV-1 WT Gag(i)mEOS2 and derivatives used in this study. (b) Western blot analysis of HIV-1 WT Gag(i)mEOS2 and mutant proteins expressed in Jurkat T cells (“cellular extract”), used for live PALM analysis, and the corresponding purified VLPs (“VLP”). Western blots are revealed with an anti-CAp24 antibody. Images of the full-length Western blots are presented in SI (Fig. S1). (c) Quantification of WT Gag(i)mEOS2 and mutant VLP production efficiency in transfected Jurkat T cells relative to total Gag protein (mean ± C.I. of 4 to 6 independent experiments). (d) Cell viability (relative to the total number of transfected cells) and protein expression of Gag(i)mEOS2 and mutants in Jurkat T cells using flow cytometry (mean ± C.I. of 3 independent experiments, see statistics section in Methods for detailed explanation). (e) Global fluorescence intensity (F.I.) of Gag(i)mEOS2 transfected Jurkat T cells as measured by flow cytometry for each condition, as indicated.