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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. 4 — Transmembrane domain-dependent ICOS-Lck association. a Surface ICOS staining of Jurkat cells stably expressing BirA*-tagged ICOS or ICOS-CD44TM molecules. b Immunoblotting of p85, Lck, and Fyn in total lysates (input) or streptavidin-precipitated proteins from Jurkat cells that stably expressed BirA*-tagged ICOS or ICOS-CD44TM molecules after 24 h of culture in the presence of exogenous biotin. The data represent the results of four independent experiments. c Input (3% total lysates) or anti-ICOS immunoprecipitates of ICOS-NT or ICOS-CD44TM-NT Jurkat cells transduced with mCherry-tagged Lck were immunoblotted for p85 and ICOS. The data represent the results of three independent experiments. d Representative TIRF-plane images of EGFP-tagged ICOS or ICOS-CD44TM (green) and mCherry-tagged Lck (red) on Jurkat cells 10 min after being placed on anti-ICOS-incorporated planar lipid bilayers. e Pearson’s correlation coefficients for ICOS-Lck or ICOS-CD44TM-Lck colocalization on Jurkat cells. Each dot denotes one cell. The data presented were pooled from two independent experiments