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. 2022 Jun 10;23(12):6516. doi: 10.3390/ijms23126516

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© 2022 by the authors.

Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).

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Physical association between CLDN4 and integrin β1 in T24 cells. (A) Effect of CLDN4 knockdown on CLDN4–ITGβ1 binding and FAK phosphorylation. (B) CLDN4 detected by immunoblotting using 0.5% Tween 20-solubilized lysate (T20) or 0.5% perfluoro-octanoic acid-soluble lysate (PFO). (C) CLDN4 bound to ITGβ1 detected by immunoprecipitation. (D) Comparison of the affinity to ITGβ1 between CLDN4 and CLDN7. Error bar indicates SD calculated by Student’s t-test from three independent trials. CLDN, claudin; BUC, bladder urothelial carcinoma; si, short interference RNA; siC, control siRNA; siCL4, siRNA for CLDN4; ITGβ1, integrin β1; FAK, focal adhesion kinase; pFAK, phosphorylated FAK. SDS-CLDN4, 0.5% sodium dodecyl sulfate-solubilized CLDN4 (whole CLDN4); T20, 0.5% Tween 20-solubilized CLDN4 (tight junction unintegrated CLDN4); PFO, 0.5% perfluoro-octanoic-solubilized CLDN4 (tight junction integrated CLDN4); ppt, immunoprecipitant.