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. 2018 Jul 18;11(5):371–382. doi: 10.1093/jmcb/mjy042

Figure 4.

Figure 4

The expression and transcriptional activation of p53 were directly induced by HIF-1α. (A and B) p53 expression in pcDNA3.1 empty vector or pcDNA3.1-HIF-1α-transfected cells (A) and pSilencer3.1 empty vector or HIF-1α-siRNA-transfected cells (B) under normoxic or hypoxic conditions for 48 h. Histograms show relative levels normalized to the loading control β-actin. #P < 0.01, *P < 0.05, **P < 0.001 compared with the pcDNA3.1 empty vector and pSilencer3.1 empty vector-transfected cells, respectively (n = 3). (C) ChIP analysis of HIF-1α binding to the p53 promoter in HK-2 cells under hypoxic condition. The reaction controls include immunoprecipitations performed using a nonspecific IgG monoclonal antibody (IgG). PCR was performed using whole-cell genomic DNA (Input). The data are representative of three independent experiments. (D) Luciferase activity of the p53 promoter reporter gene. HK-2 cells were transfected with 20 ng reporter constructs and 1 mg pcDNA3.1-HIF-1α in combination with 0.2 ng pRL-TK vector and incubated for 24 h. The luciferase activities are reported as relative light units of firefly luciferase activity normalized to Renilla luciferase activity. Data shown are mean of three independent experiments. *P < 0.01 compared with control.