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. 2018 Sep 28;9:3978. doi: 10.1038/s41467-018-06414-8

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© The Author(s) 2018

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 — Reversibility of cyclic nucleotide-dependent conformational changes. a HS-AFM time-lapse high-resolution image sequence showing CNBD conformational and 2D-lattice rearrangement dynamics of SthK upon cGMP injection (panels t = 12 min and t = 23 min) and after controlled cGMP removal and cAMP re-addition via a fluid exchange pumping system (panels t = 38 min and t = 71 min). Insets in t = 2 min, t = 23 min and t = 71 min are correlation averages (n = 22, n = 39, and n = 17 tetramers) of the corresponding frames. The molecular conformational changes lead to local membrane bending (bright area in the top part of the membrane). Scale bar: 30 nm. b cAMP (blue) and cGMP (red) concentrations as a function of time during the HS-AFM experiment. Dashed lines and adjacent labels indicate the times (bottom) and the cAMP (blue) / cGMP (red) concentrations when the images shown on top were acquired. Inset displays cGMP addition through pipetting (1) and cAMP buffer exchange (2)