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. 2022 Jul 21;609(7927):622–629. doi: 10.1038/s41586-022-05116-y

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© The Author(s) 2022

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. 4 — a–c, IL-17RB–IL-17RA interactions in the plasma membrane quantified by live-cell single-molecule imaging. a, Different receptor labelling schemes used to interrogate IL-17RB (RB) and/or IL-17RA (RA) homodimerization and heterodimerization by single-molecule co-tracking (top). Box and whisker plots show relative dimerization levels determined by single-molecule co-tracking for homodimerization and heterodimerization of IL-17RA and IL-17RB in the absence and presence of IL-25 (bottom). Numbers of cells and experiments, respectively, are 25 and 4, 49 and 4, 22 and 2, 21 and 2, 51 and 5, 48 and 5, 23 and 2, and 24 and 2 for each column, going from left to right. b,c, Heterodimerization of IL-17RA–IL-17RB (b) and homodimerization of IL-17RB (c) compared with IL-17RB mutations in the IL-17RA–IL-17RB interface. Numbers of cells and experiments, respectively, are 22 and 2, 21 and 2, 12 and 1, 13 and 1, 19 and 2, 18 and 2, 18 and 1, and 17 and 1 for each column in b, and 25 and 4, 49 and 4, 26 and 2, 29 and 2, 17 and 2, 28 and 2, 18 and 1, and 18 and 1 for each column in c, going from left to right. Statistics for a–c were performed using two-sample Kolmogorov–Smirnov tests (not significant (NS); ***P < 0.001 and ****P < 0.0001). Box and whisker plots in a–c show the five number summaries of the data: minimum, first quartile, median, third quartile, and maximum values. d–f, SPR sensorgrams reveal that IL-17RB complexes with IL-25 bind to IL-17RA. The analyte concentration range in d is a twofold serial dilution ranging from 20 to 0.156 µM. The analyte concentration ranges in e and f are twofold serial dilutions from 40 to 0.156 µM. Dissociation constants (K_d) are indicated on the sensorgrams. SPR sensorgrams represent a single experiment (n = 1) for d and one of two independent experiments (n = 2) for e and f. ND, not determined; RU, response unit. g, Model of IL-25–IL-17RB–IL-17RA signalling pathways as inferred from our structural, signalling and biophysical data.

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