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. 2024 Apr 19;22:233. doi: 10.1186/s12964-024-01612-y

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© The Author(s) 2024

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 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 — Characterization of 5-HT7R phenylalanine mutants. A. Scheme of 5-HT7R phenylalanine mutants. B, C Expression of HA-tagged 5-HT7R WT or indicated mutants in N1E-115 cells. Representative Western blot (B) and quantification (C) are shown. Data are presented as mean ± SD (N = 4, one-way ANOVA, Dunnett’s multiple comparisons. D Representative confocal images showing expression of eYFP-tagged 5-HT7R mutants. Corresponding intensity profiles are shown below. Scale bar: 10 μm. See also Additional file 12. E N1E-115 cells were transfected with cAMP FRET-based biosensor CEPAC and indicated eYFP-tagged 5-HT7R constructs. Basal cAMP levels are shown. Signals are normalized to cells transfected with 5-HT7R WT (N = 6, one-way ANOVA, Dunnett’s multiple comparisons, no statistical significance to WT). F Representative traces showing cAMP response at the single cell level after stimulation with 10 µM of 5-HT. G, H Graphs show response amplitude (G) and activation time constant (H) of cAMP relative to pretreatment (N = 6, one-way ANOVA, Dunnett’s multiple comparisons, no statistical significance to WT)