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. 2018 Jun 26;8:9706. doi: 10.1038/s41598-018-28010-y

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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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SOV inhibits the expression of HIF-1α and HIF-2α proteins and their nuclear translocation in sorafenib-resistant HCC cells. (a) HepG2-SR and Huh7-SR cells were incubated with SOV (0, 5 or 10 μM) under hypoxia (1% O₂) for 24 h. Cells were lysed and immunoblotted. Band density was normalized to β-actin. (b,c) Huh7-SR cells were incubated with SOV (0, 5 or 10 μM) under normoxia or hypoxia (1% O₂) for 24 h, and then subjected to immunoblotting (b). (c) Hypoxic Huh7-SR cells were subjected to qRT-PCR for detecting the expression of mRNAs. The level of mRNA from untreated cells was defined as 1. (d) Huh7-SR cells incubated with vehicle or SOV (10 μM) under hypoxia (1% O₂) for 24 h, and then immunostained with Abs against HIF-1α (red), HIF-2α (green) and DAPI (cellular nuclei, blue). (e) The nuclear and cytoplasmic fractions of vehicle- or SOV (10 μM)-treated hypoxic Huh7-SR cells were immunoblotted. The band density of HIF-1α or HIF-2α protein in nuclear fractions was normalized to ARNT, and that in cytoplasmic fractions, β-actin. (f) The ratio of HIF-1α or HIF-2α protein in nuclear/cytoplasmic fractions was calculated. “*” (P < 0.05) and “**” (P < 0.001) indicate a significant difference from respective vehicle-treated cells.