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. 2021 Dec 8;11:23607. doi: 10.1038/s41598-021-03013-4

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© The Author(s) 2021

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 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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(a,b) Circulating DP T cells exist as a separate population expressing naïve markers. (a) UMAP representing naïve marker genes PECAM1, TCF7, LEF1, CD7, CCR7, and SELL. (b) UMAP representing marker genes associated with thymic development. AQP3 and SMPD3 are expressed at the immature DP stage, and TOX2 and SATB1 are expressed during DP-to-SP transition. (c–f) Identification of marker gene signatures for the eight defined clusters. UMAP of paired clusters and volcano plot showing the log (fold change) and − log (p-values) for pairwise differential expression between two DP T populations. (c) CD4 CTL-like vs. Eomes⁺ Tr1-like cells. (d) Th2-like vs. Th17-like cells. (e) Tfh-like vs. Treg-like cells. (f) CD8 CTL-like vs. Innate-like cells. All plots were generated using GraphPad Prism 8.0.2. The dotted horizontal line represents a p-value of 0.05. The number in UMAP indicates the cluster number.