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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 acute experimental colitis. (A) Distal or proximal colons were excised from mice with acute colitis induced by TNBS and labeled ex vivo with 1 µM PK105b. Probe binding was detected by in-gel fluorescence (n = 3–5). *Indicates high-molecular weight species of unknown identity. (B) Immunoblotting of samples in (A) with an NE-specific antibody to reveal total NE expression. (C) Merged image of PK105b labeling (active NE; magenta) and immunoblot (total NE expression; green). (D) Immunoprecipitation of PK105b-labeled inflamed distal colon lysate with a NE-specific antibody. (E) Competition of PK105b labeling in inflamed distal colon lysates with PK101. (F) In-gel fluorescence and NE immunoblot of distal colon lysates with or without PK105b labeling. (G) Distal colon lysates from A labeled with Cy5-V-DPP.