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. 2020 Dec 4;10:21293. doi: 10.1038/s41598-020-77986-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 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/.

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Changes in pH_i do not uncouple gating from S1 motion. (A) representative inside-out PCF recording of ciHv1-I175C-TAMRA in response to repetitive voltage steps from − 80 mV to + 40 mV and back while changing pH_i and keeping pH_o = 7.0. (B) overlay of normalized mean current and fluorescence derived from the recording in (A). (C) mean fast (left) and slow (right) activation time constants of current (black) and F_signal (red) of ciHv1-I175C-TAMRA as function of pH_i while pH_o = 7 (see also Table 4). The dashed lines are linear fits with the following slopes: slope(τ_I fast) = − 0.5 log(s)/ΔpH unit, r² = 0.3, n.s.; slope(τ_{F fast}) = − 0.5 log(s)/ΔpH unit, r² = 0.4, p < 0.05; slope(τ_I slow) = − 0.67 log(s)/ΔpH unit, r² = 0.5, p < 0.05; slope(τ_{F slow}) = − 0.73 log(s)/ΔpH unit, r² = 0.5, p < 0.05; slope(τ_{I fast}) vs. slope(τ_{F fast}), n.s.; slope(τ_{I slow}) vs. slope(τ_{F slow}), n.s. (D) left, amplitude of F_signal as a function of pH_i while pH_o = 7.0 (n = 4 patches from 4 different cells). For pH_i = 6.5, F_signal = 5.0 ± 2.3; for pH_i = 7.0, F_signal = 5.8 ± 2.8; for pH_i = 7.5, F_signal = 3.4 ± 0.6; one-way ANOVA, p = 0.3. Right, baseline fluorescence at − 80 mV (F(− 80 mV)) for different pH_i while pH_o = 7.0, normalized to F(− 80 mV) for pH_i = 7.0 (n = 4 patches from 4 different cells). For pH_i = 6.5, F(− 80 mV) = 1.006 ± 0.007; for pH_i = 7.5, F(− 80 mV) = 1.001 ± 0.013; one-way ANOVA, p = 0.5. (E) representative inside-out PCF recording of ciHv1-I153C-I175C-TAMRA in response to voltage steps from − 80 to + 10 (left) or − 10 mV (right). Dashed line at the bottom indicates 0 mV. Error bars indicate SD.