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. 2017 Sep 20;7:12016. doi: 10.1038/s41598-017-12191-z

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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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HOTAIR and IGFBP2 colocalization in high-grade tumor areas (A) Whole-section, low-magnification view of an HE-stained renal-cell carcinoma containing both low-grade and high-grade tumors. (B) Immunohistochemistry (IHC) for IGFBP2, at low magnification. (C to E) Serial sections of the low-grade tumor area at high magnification, showing HE staining (C), ISH for HOTAIR (D), and IHC for IGFBP2 (E). (F to H) Serial sections of the high-grade tumor area at high magnification, showing HE staining (F), ISH for HOTAIR (G), and IHC for IGFBP2 (H). The black arrowheads indicate dot-like positive staining of HOTAIR. (I and J) Quantification of areas positive for HOTAIR (ISH, red) and IGFBP2 (IHC, brown) using ImageJ software. Total eight areas were randomly selected from low- and high-grade tumor areas. *P < 0.001.