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. 2019 Sep 16;9:13295. doi: 10.1038/s41598-019-49840-4

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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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Application of PK105b to measure NE activation in mucosal biopsies from patients with IBD. (A) Mucosal biopsies from healthy controls (n = 5) or patients with active UC (n = 9) were labeled ex vivo with 1 µM PK105b. Probe binding was assessed by in-gel fluorescence. (B) Samples in (A) were immunoblotted for total NE expression. (C) Merged image of PK105b labeling (active NE; magenta) and immunoblot (total NE expression; green). (D,E) Densitometry of active NE in (A) and total mature NE in (B), respectively. (F) Ratio of active to total mature NE in the biopsy lysates. Error bars are shown as mean ± SEM. (G) Immunoprecipitation of PK105b-labeled UC biopsy lysates with an NE-specific antibody. Gain settings for pulldown were enhanced in order to view faint bands (right panel). (H) Competition of PK105b labeling in inflamed colon lysates with PK101.