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. 2020 Jul 30;10:12824. doi: 10.1038/s41598-020-69752-y

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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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Interaction of S100A6 with tropomyosin in NIH3T3 fibroblasts. (A) Pull-down assay with the use of protein lysate from NIH3T3 fibroblasts and with CNBr-Sepharose-S100A6 affinity resin (upper panel) or CNBr-Sepharose-empty resin (lower panel). Lanes: 1—input, 2—unbound fraction, 3—last wash, 4—first wash with 250 mM NaCl, 5—last wash with 250 mM NaCl, 6—elution. Fractions were analyzed by SDS-PAGE (10% gel) followed by immunoblotting developed with mouse monoclonal anti-tropomyosin antibody, Sigma Aldrich, clone TM311. (B) Presence of S100A6–Tpm complexes in NIH3T3 fibroblasts analyzed by PLA assay. Control represents the experiment in which ligase was omitted. Complexes of examined proteins are visualized in red; cell nuclei, stained with DAPI, are in blue. Scale bar is 10 μm.