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. 2018 May 29;8:8267. doi: 10.1038/s41598-018-26636-6

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© The Author(s) 2018

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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TGFβ1 and bFGF signaling regulates Mfsd2a-dependent endothelial cell barrier functions. (A) Conditioned media from primary breast and lung metastatic cells showed significantly reduced TGFβ1 protein levels as measured using a human-specific TGFβ ELISA. (B) TGFBR2 gene expression is decreased in CD31⁺ endothelial cells sorted from freshly resected human breast brain metastasis, as compared to CD31⁺ endothelial cells fractionated from adjacent non-cancerous brain tissue, ***p < 0.001. (C) HBMECs were treated with conditioned media taken from different primary brain metastasis cells. Note the reduction in MFSD2A gene expression versus non-conditioned control media. (D) Reduced uptake of NBD-LPC by HBMECs treated with brain metastasis conditioned media, as compared to non-conditioned media, **p < 0.05 and ****p < 0.001. (E) Increased levels of VEGFR2 mRNA expression in HBMECs treated with anti-TGFβ neutralizing antibodies, as compared to treatment with species-matched control IgG, **p < 0.01. (F) Decreased NBD-LPC uptake by HBMECs treated with anti-TGFβ neutralizing antibodies as compared to cells treated with IgG control, ***p < 0.001.