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. 2020 Mar 31;24(11):3923–3937. doi: 10.1007/s00784-020-03259-8

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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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 4 — The two HA preparations significantly enhance the expression of genes encoding TGF-β1 and FGF-1 growth factors as well as the phosphorylation of Smad2 and Erk1/2 signaling molecules in ST2 and MC3T3-E1 cells. a Effect of HA1 and HA2 on Tgfb1 and Fgf1 mRNA levels in ST2 and MC3T3-E1 cells. Cells of each of the two lines were treated with each of the two HA preparations for 24 h before total RNA was extracted and analyzed by qRT-PCR. Values normalized to Gapdh are expressed relative to the values of control (Ctrl) cells. Data represent means ± SD from three independent experiments. Significant differences to the respective control, ***P < 0.001, **P < 0.01, *P < 0.05. b, c Immunoblot analyses of phospho-Smad2 (pSmad2) (b) and phospho-Erk1/2 (pErk1/2) (c) proteins in whole-cell extracts from HA-treated ST2 and MC3T3-E1 cells. Blots for total Smad2 and Erk1/2 proteins as well as the vinculin loading control are also shown. The bar charts represent densitometric quantifications of the immunoblots. pSmad2 and pErk1/2 levels are normalized to the respective total proteins used as internal controls. Data represent means ± SD from three independent experiments. Significant differences to the respective control (Ctrl) cells of each of the two cell lines, ***P < 0.001, **P < 0.01