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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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Effects of valency on initial localization and response to PKA activation. (A) Table contains the amino acid sequences of the peptides containing the different number of substrates. (B) Representative images of peptides containing one, two, and three substrates, each of which is fused to a yellow fluorescent protein (YFP), show the initial distribution (Pre) and the distribution 10 minutes post-treatment (Post) of 50 µM forskolin and 100 µM IBMX. (C) Response profile shows the normalized fluorescence signal change resulting from the activation of PKA for peptide consisting of one (blue), two (yellow), and three (black) substrates. (D) Bar chart represents the pre-treatment localization (blue) and post-treatment localization (red) as measured by taking the ratio of cytosol-to-nuclear fluorescent signal for peptide consisting of one, two, and three substrates. 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. * represents p < 0.001. Scale bar represents 10 µm.