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. 2022 Apr 5;8:30. doi: 10.1038/s41421-022-00386-2

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© The Author(s) 2022

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. 2 — a, b The display efficiencies (mean ± sd) of TCRs (868 or A6) and pMHCs (SL9-HLA-A*02:01 or TAX-HLA-A*02:01) expressed in plasmid form (a) or integrated form (b). The display efficiency was measured by FACS after staining the cells with HA-FTIC antibody at the indicated hours. Each experiment was performed with 2 technical repeats and was independently duplicated 3 times. The mean of the 6 duplicates is shown. c Comparison of the highest display efficiencies achieved among the different time points for the plasmid and the integrated forms. d The 868 TCR expressed in the integrated form recognized the SL9-HLA-A*02:01 tetramer. The SL9- HLA-A*02:01 tetramer was labeled with PE, and mCherry in MATa yeast was detected using PE-TexRed. e A schematic diagram of the yeast T cell co-culture system. The cognate T cells were constructed by stable integration of a TCR β and a TCR α gene, which were linked by the P2A linker. The pMHC-displaying yeast was the same as shown in Fig. 1a. pMHC-displaying yeasts were cocultured with cognate T cells for 20 h, followed by FACS to detect CD69 expression in T cells using an APC-conjugated CD69 antibody. f The 868 T cells and A6 T cells can be activated by cognate pMHC-displaying yeasts. Histograms showed staining by the APC-conjugated CD69 antibody. Representative data from one out of three biological replicates are shown.