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. 2020 Feb 3;11:669. doi: 10.1038/s41467-020-14526-3

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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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Fig. 3 — The experiments identify a low populated signal P3 that is in slow exchange with P2. The offset dependence of saturation for (a) the apo receptor (green), and (b) β₁AR bound to isoprenaline (red) is probed in increments of 100 Hz. c Peak intensity ratios from pairwise experiments with saturation at symmetrical offsets reveal a maximal response at −300 Hz relative to the corresponding P2 signal (grey box). The response to saturation increases from the apo form to isoprenaline bound receptor. d Saturation time course for apo receptor (green) and β₁AR bound to isoprenaline (red) with the irradiation field (field strength 25 Hz) in the on-resonance experiment (circles) applied at the position of P3, i.e. −300 Hz from P2. For the off-resonance reference experiment (crosses) saturation was applied at +300 Hz from P2. Best fits for the signal decays are shown by lines with values for k_ex and p_P3 extracted from simultaneous fits indicated for isoprenaline bound receptor as well as the apo form.