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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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Maximal effective trapping energy and diffusion of Gag(i)mEOS2 (WT and mutants) in VLPs during their assembly at the plasma membrane of Jurkat T cells. (a) Schematic representation of VLP assembly showing the different steps and their relation with localisation density (LD) changes. The final step (before VLP release) is expected to be the step were localisation density increase (LDI) and the total density are the highest. It is also expected to be the less frequent one. (b) Diagram of LDI as a function of total density (localisation density accumulation during the time of acquisition) for WT Gag, the WM and MACASP1 mutants, and NL4.3ΔPolΔEnv Gag. Red lines delimit the threshold value used for both LDI and total LD to sort the fully assembling VLPs from the others; n: in black, number of VLPs before selection, in red, fully assembling VLPs (c) Box plot of VLP assembly and release duration of the previously selected fully assembling VLP made of WM (blue), WT Gag (black) or NL4.3ΔPolΔEnv Gag (red). The average duration for particle budding was 16 ± 2 min for WM (n = 36), 20 ± 2 min for WT Gag (n = 76) and 17 ± 1.5 min for NL4.3ΔPolΔEnv Gag (n = 91).