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. 2019 Mar 5;9:3480. doi: 10.1038/s41598-019-39843-6

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© The Author(s) 2019

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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FGFR2 interacts with HIF-1α and HIF-2α in the nucleus. (a) DU145 and PC3 cells were incubated in hypoxia for 8 hours and whole cell lysates (WCL) were fractionated into cytosolic (Cyt) and nuclear (Nu) compartments. These fractions were subjected to immunoblotting with the indicated antibodies. (b) Representative immunofluorescence images. DU145 and PC3 cells were grown on coverslips and subjected to hypoxia for 8 hours. Samples were then fixed with 4% paraformaldehyde, and stained with the indicated antibodies. All samples were stained with DAPI to visualize nuclei. The scale bar represents 10 μm. (c) HEK293T were co-transfected with FGFR2-FLAG and HA-HIF-1α (or HIF-2α), and incubated under normoxic or hypoxic conditions for 8 hours. Cell lysates were subjected to immunoprecipitation with IgG, anti-FLAG, or anti-FGFR2, and the bound proteins were immunoblotted with the indicated antibodies. The blots of bound HA and bound HIF-2α were quantified using ImageJ (right panel). * denotes p < 0.05 between the indicated groups. (d) DU145 and PC3 cells were incubated under normoxic or hypoxic conditions for 8 hours. Cell lysates were subjected to immunoprecipitation with IgG or anti-FGFR2, and the bound proteins were immunoblotted with the indicated antibodies.