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. 2018 Aug 17;8:12354. doi: 10.1038/s41598-018-30799-7

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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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BMP4 and GREM1 gene expression and protein levels in unstimulated HEMFs. (a) BMP4 and GREM1 mRNA expression were measured by qRT-PCR performed on RNA isolated from primary cultures of HEMFs (HEMFs 1–4) established from normal human esophagi and from a GFP-hTERT transfected immortalized HEMF cell line (HEMF 5). Absolute and relative gene abundance are shown with delta Cts (∆Ct) wherein a higher delta Ct indicates lower gene expression. *P ≤ 0.02; **P ≤ 0.001; NS, non-significant vs. HEMF1. Values represent the mean ± SEM of three individual experiments. (b) Representative immunoblots for BMP4 and GREM1 performed on HEMFs 1–5 with β-actin as the loading control are shown. Detection of BMP4 recombinant protein as a positive control is also shown.