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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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Duplex monitoring of PIP₃ and PKA in a single cell. (A) Representative images show PIP₃ accumulation (as indicated by translocation of PH domain) and PKA activity in a single cell after EGF treatment followed by PKA activation. (B) Line-scan of the image in Fig. 4A (white line) shows the transient appearance of enriched fluorescent signal at the plasma membrane (arrows). (C) Response profile shows the normalized signal change that results from EGF stimulation followed by a saturation of the reporter response with forskolin and IBMX cocktail. Data points represent measurement from a single cell. Scale bar represents 10 µm.