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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. 3 — a The mating efficiency (mean ± sd) between the cognate TCR-pMHC pairs was significantly higher than that of the negative control pairs. The mating efficiency was calculated by the percentage of diploids detected by FACS. Seven biological repeats were measured. b A schematic diagram of the strategy used for screening cognate peptides for a given TCR in a randomly generated peptide library. The random peptide library was mated with MATa yeasts expressing the 868 TCR as the experimental group and with MATa yeasts expressing the A6 or 2 C TCR as the control group. c Ratios (mean ± sd) of the indicated SL9 mutants to the 3* peptide in diploid yeasts induced in YAMTAD screening. Each experiment was performed with 4 technical repeats and was independently duplicated 3 times. The mean of 12 duplicates is shown. d Proportion (mean ± sd) of 868 T cells activated by various SL9 mutants in the yeast T cell co-culture system. The Y-axis represents the proportion of activated T cells (CD69+) after coculturing with the indicated yeast strains. e The ROC curve depicting the FPR (false-positive rate, 1-specificity) and TPR (true positive rate, sensitivity) for library screening of TCR-pMHC interactions using the YAMTAD system. We defined a peptide as a cognate peptide if it could activate more than 10% of the T cells on average in the yeast T cell coculture assay. Unpaired two-tailed Student’s t-test was used for statistical analysis in a, c. **Indicates a two-tailed P value < 0.01. *Indicates a two-tailed P value < 0.05.