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. 2017 Jul 19;7:5824. doi: 10.1038/s41598-017-05163-w

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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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RARα agonist inhibits hepatic ApoC-III expression. (a) ApoC-III promoter activities were measured with an ApoC-III luciferase reporter assay in HepG2 cells. ApoC-III promoter reporter stable cells were generated by transfection and selection. ApoC-III luciferase reporter activities were measured after 16 hr incubation with AM580 (0.1 and 1 μM). mRNA levels of ApoC-III (b), HNF4α (c) and SHP1 (d) and protein levels of HNF4α (e) and SHP1 (f) in Hep3B cells. mRNA was analyzed by qRT-PCR in Hep3B cells after 16 hr incubation with AM580 (0.1 and 1 µM) or vehicle control. Protein levels were analyzed by western blotting with anti-HNF4α and anti-SHP1 antibodies after Hep3B cells were treated with Am580 (0.1, 1 or 10 μM) for 24 h. (g) ApoC-III protein levels are regulated by HNF4α-SHP1 axis. Hep3B cells were silenced with siHNF4α or siSHP1 in the presence or absence of AM580 (10 μM) and secreted ApoC-III levels at the end of day 3 were measured by HTRF assay as described earlier. Representative triplicate data is shown from at least three independent experiments. Liver mRNA levels of ApoC-III (h) and HNF4α (i) after 9 days of treatment with AM580 (1 mg/kg and 5 mg/kg) and vehicle control. Hepatic gene expressions were measured by qRT-PCR in liver tissue after 9 days of treatment. *P < 0.05 versus control. Data are means ± S.E. of 5–8 animals per group. *P < 0.05 versus vehicle.