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. 2022 May 3;14:46. doi: 10.1186/s13073-022-01049-3

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© The Author(s) 2022

Open AccessThis 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/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.

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Fig. 1 — Ageing leads to a marked increase in the proportion of memory-like NK2 cells among total blood lymphocytes. A t-SNE representations of the integrated single-cell transcriptomes of 11,279 PBMCs, with 4930 cells from the 4 young individuals (ages 21–28 years) and 6349 cells from the 4 elderly individuals (ages 65–68 years). Cells are coloured by cell type identity. Each dot represents a single cell. B Dot plot showing the log₂ (odds ratio) of the comparison between the cell proportions of each cell cluster in the young and elderly samples. C Scatterplot showing the logarithmic ratio between the estimated frequencies of each of the 9 cell clusters in young individuals (n = 10) and those in elderly individuals (n = 10) from bulk RNA-seq datasets (GEO103232). D Heatmap showing the top 30 differentially expressed genes for NK1 and NK2 cells from young and elderly individuals. E Violin plot showing the gene expression in NK1 and NK2 cells. F FACS staining strategy for NK1 (Lin^-CD7⁺ NKG2C⁻CD122^high) and NK2 cells (Lin⁻CD7⁺ NKG2C⁺CD122^low) from young (top) and elderly (bottom) individuals. G Bar graphs showing the proportions of NK2 cells in lymphocytes (left) and NK2 cells in NK cells (right) from young (n = 35) and elderly (n = 27) individuals. H Gene Ontology (GO) enrichment analysis of the differentially expressed genes of NK2 cells from young individuals and those from elderly individuals. The colour indicates the -log₁₀ (P-value) enrichment for each GO term. I Flow cytometry analysis of IFN-γ in NK2 cells from young and elderly individuals with or without IFN-α stimulation in vitro. J Bar graphs displaying the frequencies of IFN-γ⁺ in the NK2 subpopulation from young (n = 13) and elderly (n = 15) individuals with or without IFN-α stimulation in vitro. K Flow cytometry analysis of CD107a in NK2 cells from young and elderly individuals with or without IFN-α stimulation in vitro. L Bar graphs displaying the frequencies of CD107a⁺ in the NK2 subpopulation from young (n = 8) and elderly (n = 9) individuals with or without IFN-α stimulation in vitro. The error bars represent the standard deviation (SD). *P < 0.05, **P < 0.01, ****P < 0.0001. P-values were obtained with two-sided Student’s t-tests. The results are representative of at least three independent experiment