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. 2020 Jun 9;10:9365. doi: 10.1038/s41598-020-66349-3

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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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Basic design of the PKA kinase reporter. (A) Design consists of a NLS with three phosphorylation sites fused to a NES sequence. Phosphorylation regulates the activity of the NLS. Basic residues are colored blue; phosphoserine is colored red; and NES is colored green. (B) Response profile shows the normalized signal change resulting from the activation and inhibition of PKA FRET reporter (blue) and three variants of the nuclear-cytosol translocation reporter. The three variants are a full version with the NES (yellow), a version lacking the NES (green), and a full version with serine-to-alanine mutations (red). (C) Representative images show the changes in distribution of the full version of the reporter that occurs within three minutes of PKA activation. All data points in this figure represent an average signal intensity calculated from 30 cells over 3 independent experiments. Error bar represents standard error of mean. Scale bar represents 10 µm.