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Figure 2 | BMC Cancer

Figure 2

From: Prolyl hydroxylase 2 dependent and Von-Hippel-Lindau independent degradation of Hypoxia-inducible factor 1 and 2 alpha by selenium in clear cell renal cell carcinoma leads to tumor growth inhibition

Figure 2

Expression of HIF-α, PHD2, and PHD3 mRNA and protein in ccRCC clinical specimens and cell lines. (A) Quantitative analysis of HIF-1α, PHD2, and PHD3 mRNA by real-time RT-PCR (qRT-PCR) in ccRCC primary tumors. Expressions were normalized to the matched normal kidney tissue by calculating 2delta-deltaCT values relative to normal kidney reference. Expression of mRNA in individual tumors was shown. B. Expression analysis of HIF-1α, PHD2, and PHD3 in ccRCC cells RC2 and 786–0. Expression was normalized to endogenous β-actin by calculating delta cycle threshold (ΔCt). ΔCt = Ct value of specific gene (HIF-1α, PHD2 and PHD3) - Ct value of β-actin. The lower the ΔCt value the higher the expression of the gene. Experiment was repeated twice with triplicates and p < 0.05 was considered as significant P < 0.001. (C) Detection of HIF-1α, HIF-2α and PHD2/3 in ccRCC primary tumors and their matched normal kidney tissues by western blot analysis. 80 micrograms of protein extract was electrophoresed through Mini-Protean precast 4-20% gradient gel. Expression of β-actin was used as a loading control. (D) PHD3 protein was undetectable in ccRCC cells. Detection of HIF-α and PHD2/3 by western blot analysis in ccRCC cells RC2 and 786–0 cells. β-actin expression was used as a loading control.

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