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. 2018 Dec 12;8:17779. doi: 10.1038/s41598-018-36217-2

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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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Influence of the combinatory effect of miRNA-29b or miRNA-29b inhibitor with TGF-β1 on NHDF mRNA expression levels, XT activity and Sp1 protein expression. NHDF cell lines (n = 2) were seeded and after 24 h, transfection with a negative control miRNA (NC), miRNA-29b (miR-29b) or its inhibitor (miR-29b inh.) was performed. The cell culture medium was changed 6 h later and supplemented with TGF-β1 or vehicle (NC), as indicated. The cells were harvested 24 h after TGF-β1 supplementation for relative quantification of the mRNA expression levels of XYLT1 (a), XYLT2 (b), COL1A1 (c) and SP1 (d). Data were normalized to a normalization factor, determined by calculating the geometric mean of HPRT, GAPDH and B2M mRNA expression levels, and expressed as a ratio to one cell line. The XT activity (e) was quantified after 24 and 144 h in cell culture supernatants by using a radiochemical enzymatic assay. The XT activity in dpm was referred to the DNA content of the appropriate cell lysate. The Sp1 protein concentration (f) was measured in nuclear extracts via ELISA and referred to total nuclear protein content. Values are means ± SEM for three biological replicates per cell line *p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001 (Mann-Whitney U-test).