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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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Summary of relative cellular marker expression on CD4⁺ T cell populations using Spanning-tree Progression Analysis of Density-normalized Events (SPADE). (a) Plot showing the assigned cluster number. (b) Panel shows the cluster number, left column, and relative expression of each marker denoted as low-expression (+ white background); mid-expression (++ light grey background); mid/high-expression (+++ mid-grey background), high-expression (++++ dark grey background). Translation of SPADE cluster colour into comparative expression levels was through visual discrimination on each SPADE readout using the combined dataset. Highlighted clusters in column 1 correspond to those bearing similarities to CD4⁺ T cell populations of interest identified using t-SNE; clusters 17–18 in purple (P3), cluster 19 in red (P1) and clusters 21 and 21 in blue (P2).