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. 2018 Dec 6;17(2):143–152. doi: 10.1038/s41423-018-0183-z

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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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Fig. 1 — The ICOS transmembrane domain, but not the cytoplasmic domain, mediates the costimulation of calcium mobilization. a Surface ICOS staining of Jurkat cells lentivirally transduced with an empty vector or vectors expressing full-length ICOS or ICOS-NT. b Example of gating GFP⁺ stably transduced Jurkat cells and GFP⁻ spike-in nontransduced control Jurkat cells for the calcium mobilization assay. c Calcium mobilization in Jurkat cells of the indicated types and the respective internal GFP^- control cells after stimulation with biotinylated anti-CD3 (0.2 μg/ml) and anti-ICOS (2 μg/ml) antibodies coligated by streptavidin. The data represent the results of three independent experiments. d Surface ICOS staining of Jurkat cells lentivirally transduced with vectors expressing mutant ICOS molecules of the indicated types. e Surface ICOS staining after normalization of expression between ICOS-NT and ICOS-CD44TM-NT. f Calcium mobilization in Jurkat cells of the indicated types conducted as for c. The data represent the results of three independent experiments