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. 2020 Mar 30;18:49. doi: 10.1186/s12964-020-00537-6

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© The Author(s) 2020

Open AccessThis 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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.

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Fig. 5 — NPY displacement revealed biphasic characteristics and overexpression of Gα_i shifted receptors towards the high-affinity state. a¹²⁵I-PYY binding studies in transiently transfected Y₂R HEK293 cells after incubation with 1 μM NPY, acidic wash and receptor recovery revealed diminished binding capacity of recycled receptors (n ≥ 3). b Specific ¹²⁵I-PYY ligand displacement experiments with transiently transfected Y₂R HEK293 cells showed a biphasic binding mode with a high affinity state (black line), which is lost after the first stimulation with 1 μM NPY, followed by acidic wash and recovery in ligand free medium for 60 min (olive-green line). For normalizing the data, total binding and unspecific binding of each condition were constrained to 100 and 0% respectively. Overexpression of Gα_i shifted the percentage of the receptor towards the active state, which is summarized and represented in c). Experiments represent data n ≥ 3