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. 2016 Apr 18;8(16):25885–25896. doi: 10.18632/oncotarget.8834

Figure 4. HIF-1α transcriptionally stimulates BCL-9 expression.

Figure 4

(A) Ectopic HIF-1α expression increases BCL-9 and VEGF mRNA expression levels in SW480 and HCT116 cells. The mRNA expression levels of BCL-9 and VEGF were determined by Taqman real-time PCR and normalized with actin. (B) Ectopic HIF-2α expression increases VEGF mRNA expression levels but has no effect on mRNA BCL-9 expression levels in SW480 and HCT116 cells. (C) Ectopic HIF-1α expression increases BCL-9 protein levels in SW480 and HCT116 cells as determined by Western-blot assays. (D) and (E) HIF-1α binds to HRE-B and HRE-C sites in the BCL-9 promoter in HCT116 cells transfected with HIF-1α expression plasmids as determined by ChIP assays. The amount of DNA fragments pulled-down was determined by real-time PCR (D) or conventional PCR (E). The HRE site in the VEGF promoter serves as a positive control. (F) HIF-1α activates luciferase activity of reporter vectors containing HRE-B or HRE-C sites in the BCL-9 promoter in HCT116 cells transfected with HIF-1α expression plasmids. Luciferase reporter vectors containing the HRE site in the VEGF promoter was included as a positive control. Data are presented as mean ± SD (n = 3). *p < 0.01 (Student's t-test).