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. 2023 Nov 22;14:1221260. doi: 10.3389/fimmu.2023.1221260

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Copyright © 2023 Yang, Luan, Li, Niu, He, Zhao, Chen, Mao, Mou and Li

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Analysis of CD8+ T cells differentiation trajectories; flow cytometry and qPCR validation of Tregs and Th1-like Tregs. (A) Heatmap showing the dynamic changes of gene expression along pseudotime. (B) The trajectory of six typical genes. Unsupervised transcriptional trajectory from Monocle, colored by pseudotime (C) and cell clusters (D). (E) Representative gating strategy of flow cytometry analysis for proportion of Tregs (CD4+CD25+CD127low/-) in CD4+ T cells and Th1-like Tregs (IFN-γ+ Treg) in Tregs. (F) Flow cytometry analysis of Tregs and Th1-like Tregs. (G) Analysis of FoxP3 and CTLA-4 mRNA expression by qPCR. Data are expressed as mean ± SEM and significance was set at *p ≤ 0.05, **p ≤ 0.01.