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. 2019 Jan 24;9:540. doi: 10.1038/s41598-018-36695-4

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© The Author(s) 2019

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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HTS stimulates ex vivo ASL secretion via activation of sensory neurons and release of acetylcholine in wild-type and CF airways. (A) Treatment with lidocaine (Lido, 80 mg/ml aerosol) plus tetrodotoxin (TTX, 1 µM) (HTS, n = 45 beads from 15 tracheas; ITS, n = 49 beads from 14 tracheas; HTS + Lido + TTX, n = 18 beads from 4 tracheas; ITS + Lido + TTX, n = 9 beads from 4 tracheas). (B) Incubation with CFTRinh172 (172, 100 µM) (HTS, n = 45 beads from 15 tracheas; ITS, n = 49 beads from 14 tracheas; HTS + 172, n = 59 beads from 12 tracheas; ITS + 172, n = 29 beads from 8 tracheas). (C) Incubation with lidocaine plus TTX on CFTRinh172-treated airway (HTS + 172, n = 59 beads from 12 tracheas; HTS + 172 + Lido + TTX, n = 9 beads from 4 tracheas; ITS + 172, n = 29 beads from 8 tracheas; ITS + 172 + Lido + TTX, n = 21 beads from 6 tracheas). (D) Effect of L-703606 (NK-1 blocker, 1 µM) (HTS, n = 45 beads from 15 tracheas; ITS, n = 49 beads from 14 tracheas; HTS + NK-1 blocker, n = 36 beads from 12 tracheas; ITS + NK-1 blocker, n = 16 beads from 5 tracheas). (E) Effect of L-703606 on CFTRinh172-treated preparations (HTS + 172, n = 59 beads from 12 tracheas; HTS + 172 + NK-1 blocker, n = 10 beads from 4 tracheas; ITS + 172, n = 29 beads from 8 tracheas; ITS + 172 + NK-1 blocker, n = 17 beads from 6 tracheas). (F) Effect of atropine (1 µM) (HTS, n = 45 beads from 15 tracheas; ITS, n = 49 beads from 14 tracheas; HTS + Atropine, n = 22 beads from 5 tracheas; ITS + Atropine, n = 11 beads from 4 tracheas). (G) Effect of atropine (1 µM) on CFTRinh172-treated preparations (HTS + 172, n = 59 beads from 12 tracheas; HTS + 172 + Atropine, n = 18 beads from 5 tracheas; ITS + 172, n = 29 beads from 8 tracheas; ITS + 172 + Atropine, n = 19 beads from 5 tracheas). (H) Effect of lidocaine, TTX, and atropine on CFTR^−/− swine trachea (HTS, n = 6 beads from 2 tracheas; HTS + Lido + TTX + Atro, n = 4 beads from 2 tracheas; ITS, n = 4 beads from 2 tracheas). Data are presented as mean ± SEM and values at 18 min were analyzed with ANOVA and Tukey’s multiple test. Data sets labeled with different letters differ significantly, p < 0.05.