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. 2020 May 15;11:2440. doi: 10.1038/s41467-020-16299-1

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© The Author(s) 2020

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. 2 — a The IN/IC formats allow splicing between bait and prey. In the CIC orientation, IC-FLAG is fused to the C-terminus of a prey protein (Prey-IC-FLAG). Its combination with IN bait (Bait-V5-IN) leads to the transfer of FLAG tag to the bait generating Bait-V5-FLAG. In the NIN orientation, V5-IN is fused to the N-terminus of a bait (V5-IN-Bait). Its use with IC prey (IC-FLAG-Prey) causes the transfer of V5 tag to the prey thus generating V5-FLAG-Prey. The CIC-GFP construct (Prey-IC-FLAG-GFP) is created to allow the detection of NIN/CIC-GFP combination, which produces a V5-FLAG-GFP peptide. b The performance of different SIMPL formats was experimentally assessed using the rapamycin-induced FRB/FKBP1A interaction in which the corresponding bait and prey constructs were transiently transfected. Bands of spliced products are highlighted with triangles and parental proteins are highlighted with asterisks. The densities of spliced bands (FRB-FKBP1A) were quantified with ImageJ and are presented as bar graphs above the blots. The blot is representative of three independent experiments. Source data are available in the Source Data file.