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. 2017 Oct 4;7:12679. doi: 10.1038/s41598-017-12976-2

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© The Author(s) 2017

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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Effect of 991 and C13 on BMP6-induced hepcidin gene expression in WT and AMPKLiv KO mice. (A) Immunoblots showing pAMPK/AMPK and pACC/ACC in hepatocyte extracts prepared as previously described, treated with 10 nM BMP6, 3 μM 991 or 3 μM C13. (B) Q-PCR analysis showing hepcidin mRNA levels in mouse primary hepatocytes treated with BMP6 (10 nM), 991 (3μM) or C13 (3μM) for 24 h. RNA extraction and real-time quantification of transcripts were performed as described. mRNA expression was calculated using the ΔΔCt method and normalized to the expression of cyclophilin A. Hepcidin gene expression is expressed in % of BMP6 treated hepatocytes. Error bars represent SD for n = 3 samples in each group.