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. 2020 Apr 3;10:5852. doi: 10.1038/s41598-020-62615-6

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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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Opposite charges at position 116 (Na_VSp1 S4-S5_L) and 222 (Na_VSp1 S6_T) stabilizes the Na_VSp1 channel open state. (A) representative, superimposed current recordings of single Na_VSp1 mutant D222R and double mutant D222R/R116D. Activation voltage protocol used is the same as in Fig. 2. (B) Dot plot and mean ± sem of peak current densities recorded in COS-7 cells transfected with D222R or D222R/R116D Na_VSp1, at 30 mV. (C) Relative peak conductance versus membrane potential curves for D222R or D222R/R116D Na_VSp1 channels transfected in COS-7 cells. Lines are Boltzmann fits to the data. (D,E) Dot plot and mean ± sem of Na_VSp1 half-activation potential (V_0.5; D) and activation slope (K; E) in COS-7 cells transfected with D222R or D222R/R116D Na_VSp1. ***p value vs. D222R < 0.001.