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. 2018 May 22;8:8003. doi: 10.1038/s41598-018-26172-3

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© The Author(s) 2018

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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Single cell analysis of time-lapse images reveals that CD4⁺ T cells exhibit distinct intracellular calcium signatures when interacting with sAgs-pulsed MDMs. (A and B) Intracellular calcium profiles of CD4⁺ T cells recorded with a 340:380 ratio exceeding the threshold value (≥1); plots show the intracellular calcium fluxes of CD4⁺ T cells over time, when perfused over untreated (A) or sAgs-pulsed (B) MDMs; the MDM (large white cell) interacting CD4⁺ T cell considered in each plot is visible in the adjacent image frames, highlighted with a red track and labeled according to the 340:380 color-scale. (C) Calcium signatures of CD4⁺ T cells on MDMs were grouped into four categories according to their 340:380 ratio profiles, as illustrated with representative plots. (D) Charts indicating the percentage of CD4⁺ T cell calcium signatures classified into each group in the presence and absence of sAgs. Scale bars = 20 µm.