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. 2020 Mar 10;10:4446. doi: 10.1038/s41598-020-61441-0

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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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CXCR2 knockdown via siRNA transfection in brain endothelial bEnd.3 cells resulted in reduced activation of the downstream angiogenic pathway. Brain endothelial bEnd.3 cells cultured in DMEM for 24 h were transfected with siRNA targeting CXCR2 (90 pmol) and incubated for 24 h. (a) CXCR2 knockdown resulted in the downregulation of IL-8-downstream p-p38, angiogenic growth factors, IκB degradation representing NFκB release, and CD31 (endothelial marker) according to Western blotting, despite IL-8 treatment. (b–i) The graphs depict the band intensity in each Western blot assay. Data are shown as the mean ± SEM (each n ≥ 3). The data were obtained from triplicate results. Asterisks (*) indicate significant differences (*p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001, one-way ANOVA). IL-8, bEnd.3 cells treated with IL-8 without prior CXCR2 knockdown; siCXCR2+IL-8, CXCR2-knockdown bEnd.3 cells treated with IL-8.