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. 2020 Aug 25;10:14151. doi: 10.1038/s41598-020-70822-4

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© The Author(s) 2020

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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TRPV4 colocalizes and interacts with cytoskeletal protein F-actin. (A) U87 cells were plated on dishes for IF, and F-actin was labeled with FITC-phalloidin (green). TRPV4 was labeled with anti-TRPV4 antibody followed by TRITIC-conjugated antibody (red). Inset images show the TRPV4 distribution at the membrane and cellular protrusions. The colocalization profiles of inset images were analyzed by ImageJ and are listed below the image. Scale bar = 20 µm and 2 µm. (B) GL261 cells on confocal dishes for F-actin and TRPV4 staining to show colocalization. The inset images and colocalization profiles are listed below. Scale bar = 10 µm and 1 µm. (C-D) Coimmunoprecipitation of TRPV4 and F-actin: the whole-cell lysate was immunoprecipitated (IP) with anti-TRPV4 or F-actin antibody, and the immunocomplexes were immunoblotted (IB) with the opposite antibody in U87 and GL261 cells.