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. 2019 Nov 11;9:16461. doi: 10.1038/s41598-019-52839-6

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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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Contribution of NFκB to the ET-1-induced oxidative stress and COX-2 expression in VSMC from hypertensive rats. Effect of lactacystin (Lac, 10 µM) on NOX-1 mRNA levels (a), NADPH oxidase activity (b), COX-2 protein expression (c) and mRNA levels (d) induced by ET-1 (0.1 µM, 1 h). (e) Effect of ET-1 (0.1 µM, 45 min) on nuclear p65 NFκB protein expression in VSMC from SHR; a representative blot of the cytosolic (Cy) and nuclear (Nu) expression is shown below; TATA-binding protein (TBP) and GAPDH (after reblotting) cytosolic and nuclear expressions are also shown to guarantee the successful cellular fractioning. (f) Representative photomicrographs of p65 NFκB immunofluorescence (red) in VSMC from SHR in control and after incubation with ET-1 (0.1 µM, 45 min, n = 7). Negative controls without primary or secondary antibody are also shown. Bar scale represents 50 µm. Statistical analysis by Student’s t-test; *P < 0.05 vs control and ^#P < 0.05 vs ET-1. n denotes number of experiments. Full-length blots are presented in Supplementary Fig. S3.