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. 2019 Jun 27;294(33):12507–12520. doi: 10.1074/jbc.RA119.008255

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© 2019 Wichmann et al.

Published under exclusive license by The American Society for Biochemistry and Molecular Biology, Inc.

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Figure 2. — Extracellular pH affects gating of Xenopus-δβγ ENaC. a, representative current traces from cell-attached patch-clamp recordings of oocytes expressing Xenopus δβγ-ENaC at a holding potential of −100 mV. Recordings were performed using pipette solutions at pH 8.0, 7.4, or 6.0 (pH_pip). Dashed lines indicate the number of individual open channel levels or the current baseline (c). b, open probability of individual channels increases with decreasing pH_pip (one-way ANOVA, F = 24.93, p < 0.0001; Tukey's multiple comparisons test). c, number of visible channels in cell-attached recordings as depicted in a (one-way ANOVA, F = 5.83, p = 0.0056; Tukey's multiple comparisons test). d, current (I)/voltage (V_hold) plots derived from cell-attached patch-clamp recordings of Xenopus δβγ-ENaC at different pH_pip. The pH_pip does not affect the channel's slope conductance (G_slope) (one-way ANOVA, F = 1.854, p = 0.1725), which was calculated from the linear regression of unitary channel conductance (mean of at least three single channel amplitudes per n) at −40 to −100 mV. e, estimation of the number of channels in the patch. P_n is the probability of n channels out of the total number of channels within the patch being opened. The observed probability for each n (i.e. observed current levels) was compared with a theoretical distribution of P_n as predicted by a binomial distribution, assuming n channels present in the patch. There is no significant difference between observed and predicted P_n values under the employed pH_pip conditions (Kruskal-Wallis test with Dunn's multiple comparisons test in each panel. p > 0.9999 between each pair of P_n observed and P_n predicted.). Statistical evaluation is based on individual recordings lasting for 120–180 s with a maximum of eight channels per patch. Please note that single-channel characteristics at pH_pip 7.4 include data, which have been reported earlier (18). Patch-clamp data for both studies were collected simultaneously using the same oocyte batches for all pH conditions and αβγ- as well as δβγ-ENaC.