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. 2021 Mar 19;28(8):2436–2449. doi: 10.1038/s41418-021-00761-8

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© The Author(s), under exclusive licence to ADMC Associazione Differenziamento e Morte Cellulare 2021

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 HeLa (averaged traces ± SD of six cells from one experiment) or B HT-29 cells (averaged traces ± SD of seven cells from one experiment) expressing the cAMP-sensitive FRET-based sensor H187 were challenged with increasing doses of forskolin (FSK) followed by FSK (20 µM) together with IBMX 500 µM (FSK/IBMX) to saturate the sensor. C HeLa cells expressing the PKA-dependent phosphorylation sensor AKAR4 marginally responded to FSK alone; however, addition of IBMX 500 µM produced a near-saturation response. Saturation was achieved by FSK/IBMX (averaged traces ± SD of 20 cells from four independent experiments). D In HT-29 cells AKAR4 gave extremely low responses independent of the treatment (averaged traces ± SD of 37 cells from 15 independent experiments).