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. 2020 Jun 29;11:3272. doi: 10.1038/s41467-020-17046-2

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© The Author(s) 2020

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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Fig. 5 — Next-generation sequencing-based high-throughput TCR-β CDR3 analysis of bulk sorted T-memory CD4+ Tconvs and Tregs from matched NI and I TDLNs and the primary tumor. Graphs a–d display the TCR-β CDR3 clones of memory CD4+ Tconv a, b; or effector Tregs c, d. Graphs a, c show cumulative frequencies of from NI TDLNs (blue), I TDLNs (red), and the corresponding primary tumor (green), for two representative patients. Stacked bar chart b, d depict the usage frequencies of the top 100 tumor TCR-β CDR3 sequences and their distribution in matched NI and I TDLNs for two representative patients. Numbers in the graphs indicate percentage of shown TCR-β CDR3 sequences out of total TCR-β CDR3 sequences in the sample. e, f TCR repertoire overlap between Tregs and Tconvs. e Shown is the number (%) of unique TCRs shared between Tregs and Tconvs among all Treg clones in TDLN (N = 5) and tumor (N = 2). f Shown is the number (%) of unique TCRs shared between all tumor Tregs and Tconvs present in the NI or I TDLNs (N = 2). Each symbol is a sample, and samples from the same patients are in the same color.