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. 2020 May 26;10:8744. doi: 10.1038/s41598-020-65761-z

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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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IP₃ does not permeate channels made of Cx50. Three different Cx50 cell pairs (A,C,E) all showed a rapid spike in Ca²⁺ fluorescent intensity in cell 1 (red lines) when IP₃ was introduced. In contrast to Cx43, cell 2 (green lines) never showed any spike in fluorescent intensity (B,D,E) for up to 220 seconds following IP₃ delivery to cell 1, indicating no permeation through Cx50 channels. When 500 µM IP₃ was directly introduced into cell 2 after monitoring fluorescence for 200 seconds following IP₃ release into cell 1 (E,F), a rapid spike in fluorescent intensity was observed in cell 2 within seconds. The mean gap junctional conductance (± SD) for the three Cx50 cell pairs shown was 23 ± 2.6 nS.