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. 2017 Jun 14;7:3517. doi: 10.1038/s41598-017-03742-5

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© The Author(s) 2017

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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H₂S targets phosphodiesterase rather than adenylyl cyclase. (a) Representative current traces of TEVC recordings of CFTR-expressing oocytes. Transmembrane currents (I_M) were recorded and oocytes were either exposed twice to forskolin (fsk., 5 µM; black bars; left trace) or first to forskolin and then to a combination of forskolin and 50 µM Na₂S (grey bar; right trace). (b) Statistical analysis of data obtained from experiments as shown in panel a. Depicted are normalised values from individual experiments (grey symbols) as well as means ± SEM of forskolin (fsk.) effects. This represents the ratio of the first and second current stimulated by forskolin (***P ≤ 0.001, Mann-Whitney test). (c) Representative current traces of TEVC recordings of CFTR-expressing oocytes. Oocytes were either exposed twice to IBMX (1 mM; black bars; left trace) or first to IBMX and then to a combination of IBMX and 50 µM Na₂S (grey bar; right trace). (d) Statistical analysis of data obtained from experiments as shown in panel c. Depicted are normalised values from individual experiments (grey symbols) as well as means ± SEM of IBMX effects. This represents the ratio of the first and second current stimulated by IBMX Student’s unpaired t-test with Welch’s correction). (e, f) Representative current traces of TEVC recordings of CFTR-expressing oocytes. (f) Oocytes were exposed twice to membrane-permeable 8Br-cAMP (100 µM, grey bars) in the presence of the AC-inhibitor MDL 12330 A (MDL, 20 µM; black bars). The perfusion was stopped (indicated by the number symbol’) when 8-Br-cAMP was in the perfusion chambers in order to avoid massive consumption of this compound. Perfusion was started again at the time of drug removal. (e) The same protocol was employed, with the exception that Na₂S (50 µM, black arrowhead) was applied before the second exposure to 8Br-cAMP. (g) Statistical analysis of data obtained from experiments as shown in panels e and f. Depicted are values of the first (1) and second (2) 8-Br-cAMP-induced current (I_cAMP) from individual experiments (grey symbols) as well as means ± SEM (**P ≤ 0.01, Student’s paired t-test). Numbers of experiments (n) are indicated in parentheses.