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. 2019 May 22;9:7679. doi: 10.1038/s41598-019-44194-3

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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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Effect of downregulated MYH9 expression on the F-actin network. (A) Validation of siRNA knockdown of Myh9 (siMYH9) in podocytes using real-time PCR, corrected by β-actin mRNA levels in the same sample. (B) Western blots of MYH9 after siRNA-mediated inhibition of MYH9 in control (Con) and Ang II-treated podocytes with or without losartan (Los). The blots were cropped from different parts of the same gel. (C) Cultured podocytes grown on coverslips were fixed with 4% PFA and immunolabeled with FITC-phalloidin (green) and junctional marker ZO-1 (red). Control cells display uniformed actin stress fibers and ZO-1. Treatment of podocytes with Ang II resulted in actin rearrangement and loss of ZO-1 staining. MYH9-depleted cells showed disorganized, shortened and decreased stress fibers. Treatment of Ang II-stimulated control or MYH9-depleted cells with losartan restored actin stress fibers and ZO-1 staining. (D) podocytes transfected with GFP-MYH9 were analyzed by immunofluorescence with FITC-phalloidin and ZO-1 antibodies. Magnification 40x; bar = 50 μm. Data are presented as the means ± SD, n = 3. Similar results were obtained in three independent experiments. *P < 0.05 compared with the siCTRL. ^#P < 0.05 compared with siCTRL and siMYH9 control podocytes treated with Ang II. ^†P < 0.05 compared with the siMYH9 control. Abbreviations: siCTRL, scrambled control; siMYH9, MYH9 siRNA transfection.