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. 2018 Oct 2;8:14675. doi: 10.1038/s41598-018-32821-4

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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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Depolarization of SH-SY5Y cells increases the interaction between Arc-mCherry and EGFP-PICK1. (a) The amount of Arc/PICK1 complex formation is increased under depolarizing conditions based on the G(0) values from the cross-correlation functions obtained from RICS (ccRICS) analysis. (b,c) Intensity images of EGFP-PICK1 in non-depolarizing and depolarizing buffer (respectively) show areas of PICK1 localization. (d) Phasor points of a co-transfected cell under depolarizing conditions show a population of unquenched pixels (no interaction) circled in green with a lifetime of 2.3 ns and a distinct second population of quenched pixels (interaction between Arc-mCherry and EGFP-PICK1) circled in red with a much lower lifetime of 1.8 ns. (e,f) Spatial arrangement of phasor points on representative non-depolarized and depolarized cells (respectively) indicate the location of pixels that do not show FRET (e; green) and pixels showing FRET (f; red).