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. 2017 Nov 29;7:16552. doi: 10.1038/s41598-017-16457-4

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© The Author(s) 2017

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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Association of DHHC5 with S1P₁R. (a) SH-SY5Y cells transiently expressing S1P₁R-CFP and either YFP-DHHC5 or YFP-DHHC20 were fixed and observed for each fluorescence by confocal laser scanning microscope. (b) Cells expressing various proteins as in (a) were fixed for FRET efficiencies. Values represent means ± s.e.m. of 3 independent experiments carried out in triplicate. Statistical significance was analysed by Student’s t-test (**P < 0.01 versus DHHC5-expressing cells). (c) Cells (2 × 10⁷ cells) expressing FLAG-S1P₁R were lysed and immunoprecipitated by FLAG-antibody. The immunoprecipitates or lysates were subjected to immunoblot analysis using an anti-DHHC5 or an anti-S1P₁R antibody. Representative images form 3 independent experiments are shown. Full-length images are presented in Supplementary Figure 7.