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. 2017 Nov 9;7:15140. doi: 10.1038/s41598-017-15474-7

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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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Hypoxia increases intracellular ROS levels. (A) Representative cropped immunoblot (left) and densitometry analysis (right) of HIF1α protein levels in MDA-MB-468 cells exposed to hypoxia (6 h) compared to normoxic cells (full-lenght immunoblot is shown in the Supplementary Fig. S2). (B) CA9 mRNA levels in normoxic and hypoxic (24 h) cells were assessed using real time RT-PCR. (C) Intracellular levels of ROS measured by DCF-DA assay in cells exposed to hypoxia (12 h) compared to control cells (remained in normoxia). Graphs represent the mean ± SD for three independent experiments. *p < 0.05, **p < 0.01 (unpaired t-test).