Figure - PMC

Skip to main content

An official website of the United States government

Here's how you know

Here's how you know

Official websites use .gov
A .gov website belongs to an official government organization in the United States.

Secure .gov websites use HTTPS
A lock ( ) or https:// means you've safely connected to the .gov website. Share sensitive information only on official, secure websites.

View full-text article in PMC

. 2017 Nov 6;7:14514. doi: 10.1038/s41598-017-15108-y

Search in PMC
Search in PubMed
View in NLM Catalog
Add to search

© 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/.

PMC Copyright notice

TR-FRET assay for detecting interaction between claudin-4 and C-CPE. (a) Schematic representation of the TR-FRET-based assay for detecting the interaction between claudin-4 and C-CPE. When claudin-4 binder binds to the site of claudin-4 and C-CPE interaction, FRET signal is not detected. (b) Validation of the TR-FRET-based claudin-4 and C-CPE binding assay. The assay was performed by sequentially adding europium (K)-anti-His antibody (Ab), His-tagged claudin-4 (His-claudin-4) and XL665-anti-GST Ab, followed by the addition of GST-tagged C-CPE or C-CPE Y306A/L315A. TR-FRET signals were measured with an Artemis HTRF plate reader at 620 and 665 nm emission wavelengths. Data are means ± SD (n = 3). (c) Inhibitory effect of claudin-4 binders on the interaction between His-claudin-4 and GST-C-CPE in the TR-FRET-based assay. C-CPE, C-CPE Y306A/L315A or anti-claudin-4 Ab (either against the extracellular domain [ex] or the intracellular domain [in] of claudin 4) was incubated with His-claudin-4, and then GST-C-CPE, XL665-anti GST Ab and Eu(K)-anti His Ab were sequentially added. Data are means ± SD (n = 3).