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. 2019 Oct 30;9:15650. doi: 10.1038/s41598-019-51961-9

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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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Spanning-tree Progression Analysis of Density-normalized Events (SPADE) of CD4⁺ T cells. (a) Representative gating strategy of live CD4⁺ T cells from one individual. (b) SPADE analysis of live CD3⁺CD4⁺ cells showing combined flow cytometry standard (FCS) dataset from all participants at all time-points. Cluster size indicates relative frequency of live CD3⁺CD4⁺ events. Cluster colour indicates relative marker expression; with scale from green (low) to red (high).