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. 2018 Apr 10;9:1354. doi: 10.1038/s41467-018-03728-5

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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. 5 — Switch of reversal potentials between co-expressed rASIC3 and rP2X3Rs in CHO cells. Whole-cell patch-clamp recordings at a holding potential of −65 mV. a Current traces induced by protons (pH 6.5) and α,β-meATP (10 µM) applied individually onto CHO-rASIC3 (left panel) and CHO-rP2X3R cells (middle panel) or successively onto CHO-rASIC3/P2X3 cells (right panel). Agonist application was at holding potentials which increased stepwise from −60 to + 90 mV in 30 mV increments. Current–voltage relationships were constructed from recordings similar to those shown in (a) in order to determine the reversal potentials (E_rev) (b). Means ± S.E.M. of the indicated number of experiments. c Measurement of E_rev of protons (left panel) and of α,β-meATP (right panel) by a similar protocol as shown in (a) on separate populations of CHO-rASIC3/rP2X3R cells. Under these conditions, the E_rev values of these two agonists were interchanged. Current–voltage relationships constructed from recordings similar to those shown in (c) in order to determine the E_rev (d). Means ± S.E.M. of the indicated number of experiments. Blockade of ASIC3 channel activity by the selective antagonist APETx2 (0.1 µM) shifted the E_rev of α,β-meATP back near to its original value of around 0 mV. e In CHO-rASIC3/rP2X3R cells, ramps of 200 mV duration (−90 to +90 mV) were delivered at a holding potential of −65 mV to determine the E_rev of ASIC3 after a drop of the normal pH from 7.4 to 6.5. Then, a high concentration of α,β-meATP (100 µM) was applied in order to change the distribution of extra- and intracellular ions. Eventually, the E_rev was re-determined after washing out α,β-meATP. Original tracing; large capacitive artifacts at the beginning and end of the ramp-induced currents were retouched. f Current–voltage curves were constructed from the indicated number of experiments similar to that shown in (e). The scale labels for the vertical bars were 2 nA (a, left panel), 1 nA (a, middle and right panels), 1 nA (c) and 500 pA (e). The scale labels for the horizontal bars were 1 s (a, c) and 2 s (e)