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. 2017 Aug 8;7:7546. doi: 10.1038/s41598-017-07973-4

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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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Upregulation of HIF-1α in human PCa. (a) HIF-1α protein was detected by western blot in nonmalignant (RWPE-1 and BPH1) and PCa cell lines (PC-3, DU145, LNCaP, and 22RV1) as indicated. (b–d) Total RNA extracted from RWPE-1, BPH1, PC-3, DU145, LNCaP, and 22RV1 cells was subjected to qRT-PCR for HIF-1α (b), VEGF (c) and GLUT4 (d). (e) The HIF-1α promoter-driven reporter (firefly luciferase) and a control vector (Renilla luciferase) were co-transfected into RWPE-1, BPH1, PC-3, DU145, LNCaP, and 22RV1 cells for measurement of luciferase activity. HIF-1α promoter activity was calculated as a ratio of firefly to Renilla activity. (f) Human normal and malignant tissue (Gleason score 9) sections were probed with HIF-1α antibody (scale bars, 100 µm). (g) Protein expression of HIF-1α, VEGF, and GLUT4 were examined with western blot, in PC-3, DU145, and LNCaP cells after various treatments as indicated. Data are expressed as mean ± SD of seven independent experiments. ^$p < 0.05 versus RWPE-1 or BPH1 cells or normal tissue. *p < 0.05 versus control group. ^#p < 0.05 versus si-HIF-1α or DDP group. Original blots are shown in Supplementary Figure 5. C: Ctrl; D: DDP; S: si-HIF-1α; D/S: DDP/si-HIF-1α.