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. 2022 Jul 13;13:4046. doi: 10.1038/s41467-022-31519-6

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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. 4 — a 2-D dot plots showing the expression of selected genes in the different CD4⁺ T-cell clusters in ACPA+ and ACPA− SF (circle size indicates the percentage of cells expressing, color intensity indicates average expression) (n = 4 ACPA+, n = 4 ACPA− RA SF). b Representative flow cytometry dot plots showing the expression of LAG-3, CXCR6, CD69, CD49a, and CX3CR1 on GPR56⁺ and GPR56⁻ CD4⁺ T cells in ACPA+ RA, quantified in ACPA+ and ACPA− SF in c) P < 0.0001 (%LAG-3), P = 0.0052 (%CXCR6), P < 0.0001 (%CD69), P < 0.0001 (%CD49a), P = 0.0379 (%CX3CR1) in GPR56⁻ versus GPR56⁺ in ACPA+ SF; P = 0.0002 (%LAG-3), P = 0.0006 (%CXCR6), P = 0.0078 (%CD69), ns (%CD49a), P = 0.0262 (%CX3CR1) in GPR56⁻ versus GPR56⁺ in ACPA− SF; P < 0.0001 (%LAG-3) in GPR56⁺ in ACPA− versus ACPA+; P = 0.0052 (MFI LAG-3), P = 0.0007 (MFI CXCR6), P = 0.0002 (MFI CD69), P = 0.0196 (MFI CD49a) in GPR56⁻ versus GPR56⁺ in ACPA+ SF; ns (MFI LAG-3), P = 0.0002 (MFI CXCR6), P = 0.0002 (MFI CD69), ns (MFI CD49a) in GPR56⁻ versus GPR56⁺ in ACPA − SF; (n = 10 ACPA+, n = 8 ACPA−, LAG-3, CXCR6, CD69, CD49a) (n = 8 ACPA+, n = 7 ACPA−, CX3CR1). White dots indicate GPR56⁻ CD4⁺ T cells and black dots indicate GPR56⁺ CD4⁺ T cells. d Expression of LAG-3, CXCR6, CD69, CD49a and CX3CR1 on CD4⁺ T cells in SF; P = 0.0005 (%LAG-3 in ACPA− versus ACPA+); (n = 8 ACPA−, n = 10 ACPA+, LAG-3, CXCR6, CD69, CD49a) and (n = 7 ACPA−, n = 8 ACPA+, CXC3CR1). Blue dots indicate ACPA− RA SF and red dots indicate ACPA+ RA SF. c, d Line represents median, two-tailed Mann–Whitney U test. Data are from a pool of eight independent experiments where a circle is a single replicate. e Correlation between the frequency of LAG-3 in CD4⁺ T cells in RA SF and the levels of anti-CCP (cyclic citrullinated peptide) antibodies (n = 18), P < 0.0001, Spearman two-tailed test. ns not significant.