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. 2022 Jan 10;13:181. doi: 10.1038/s41467-021-27801-8

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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 UMAP dimensionality reduction embedding of PBMCs from DFU-Healers, DFU-Non-healers, Healthy subjects, and non-DFU DM patients. The identified cell types were DCs: dendritic cells; VasEndo: vascular endothelial cells; T-lympho: T lymphocytes; CD8T1: CD8⁺ T lymphocytes cluster 1; CD8T2: CD8⁺ T lymphocytes cluster 2; NK: natural killer cells; NKT: natural killer and T cells; B-lympho: B lymphocytes; CD14Mono: CD14⁺ monocytes; CD16Mono: CD16⁺ monocytes. Dotted lines are drawn around cell groups of similar lineages. b Bar plots showing percentage of T-lymphocytes (T-lympho) and CD8⁺ T lymphocytes cluster 2 (CD8T2) per percentage of NKT cells in the CD45⁺ subset of cells across various clinical groups. DFU-healers depict significantly higher ratio of T-lympho and CDT2 cell cluster in comparison to DFU-Non-healers and Diabetic. Data represent the mean and standard error of mean (SEM) values from n = 2 Non-Healers, n = 3 Healers, n = 2 Diabetic and n = 3 Healthy subjects. p = 0.01 for Healers vs Non-Healers and p = 0.006 for Healers vs Diabetic in T-Lympho; p = 0.036 for Healers vs Non-Healers and p = 0.035 for Healers vs Diabetic in CD8T2 using two-sided Welch’s t-test. c Heatmap showing significant DEGs in Healers compared to Non-healers in the T-lympho, CD8T2 and NKT cell clusters. d Biological pathways that are significantly (P value < 0.01) activated (Z score >1.5) /inhibited (Z score < −1.5) in T-lympho, CD8T2 cells of Healers in contrast to NKT cells of Non-healers. Activation and inhibition of key upstream regulators is shown in pseudo color, where blue represents inhibition, and red represents activation. e Upstream regulatory molecules significantly inhibited (blue) in the T-lympho and CD8T2 cells of Healers as compared to Non-healers at the systemic level. Legend shows shapes and lines annotation for the regulatory network. f Violin plots showing expression levels of 3 key upstream regulator molecules NFKBIA, CCL5, and TGFB1, in the NKT, T-Lympho, and CD8T2 clusters.