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. 2019 Jan 24;9:636. doi: 10.1038/s41598-018-37213-2

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© The Author(s) 2019

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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Development of split luciferase probes for detecting RAF dimerization. (a–d) Identification of the best probe pairs for detecting RAF dimerization using different orientations of RAF and ELuc fragments. 293T cells were transiently co-transfected with the indicated probe pairs (a), stimulated with 1 µM LY3009120 or the vehicle control (0.1% DMSO) for 2 hours, and luciferase activities were measured (b–d). (e–g) Optimization of the linker length. 293T cells were transiently co-transfected with the indicated probe pairs, stimulated with 1 µM LY3009120 or the vehicle control (0.1% DMSO) for 2 hours, and luciferase activities were measured. Results are presented as relative luminescence units (RLUs) (means ± SD, n = 3) and the ratio of RLUs measured in LY3009120-treated cells to DMSO-treated cells.