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. 2017 Jul 26;7:6623. doi: 10.1038/s41598-017-06162-7

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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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Rap2a enhances RCC cell metastasis in vivo. (A) 786-O cells were examined for their infection efficiency under the fluorescent microscope after lentivirus transfection (×100 magnification) (top panel). Western blot of Rap2a from Rap2a-overexpression 786-O cell lines and Ctrl-786-O cell lines selected with puromycin for 3 weeks after lentivirus infection. Rap2a expression levels remain stable without puromycin selection for 2 months (bottom panel). (B) Representative images of 10% buffered formalin fixed lungs with metastatic nodules 2 months after respective injection of Ctrl, Rap2a-overexpression 786-O cell lines. Arrows indicate metastatic nodules. The number of lung metastatic nodules was counted under a dissecting microscope. (C) A statistically dramatic increase in the number of the lung metastases was seen in Rap2a-overexpression group compared with the Ctrl group. Data are displayed with means ± SD from 10 mice in each group. (D) Immunostaining of Rap2a and p-Akt in metastatic nodules of Rap2a-overexpression and Ctrl 786-O groups. Rap2a and p-Akt expression in Rap2a-overexpression group were much higher compared with Ctrl group. **P < 0.01.