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. 2023 Jun 21;13:10082. doi: 10.1038/s41598-023-36468-8

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© The Author(s) 2023

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 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/.

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Kinetics of PD-1 and LAG-3 surface expression following a multi-hour α-GalCer iNKT cell activation and rest assay (n = 4). (A) Example of iNKT flow cytometry gating strategy. (B) LAG-3 proportion of parent population expression (left y-axis, red line), (C) PD-1 proportion of parent population expression (left y-axis, red line) and (D) median fluorescent intensity (MFI) of PD-1 + iNKT cells (left y-axis, red line) in relation to proportion of parent population IFN-γ production (right y-axis, blue line) over the multi-hour α-GalCer stimulation time course. Mean and 95% confidence intervals are represented at each timepoint for both datasets, with the mean represented as either circles (left y-axis) or squares (right y-axis). Raw data values are represented as smaller open dots. Paired two-tailed T-tests have been selected to highlight specific dataset trends (dashed blue line: right y-axis dataset; straight red line: left y-axis dataset), where p < 0.05 were considered significant (*< 0.05, **< 0.01, ***< 0.001, ns: not significant).