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. 2018 Jun 12;8:8984. doi: 10.1038/s41598-018-27174-x

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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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Compatibility with optogenetic tools. (a,e) Scheme of light-induced activation of ERK or S6K. HeLa cells expressing hyBRET-ERKnls and mCherry-CRY2-cRaf (b–d) or hyBRET-S6K and mCherry-Cry2-iSH2 (f–h) were illuminated with 490 nm light for 100 ms. The BRET image in IMD mode and mCherry fluorescence image are shown in (b,f). (c,g) The cytoplasmic intensity of mCherry was normalized to the mean value before stimulation and used as a translocation index. (d,h) The BRET ratio normalized to the mean value before stimulation was used as normalized BRET. Averages of single cells are shown with the mean value (red). Bars, 10 μm.