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. 2014 Feb 5;9(2):e88301. doi: 10.1371/journal.pone.0088301

Figure 2. PIM2 is a HIF-1α target gene.

Figure 2

(A) Schematic representation of the hypoxic response element (HRE) binding sites in the PIM2 promoter region. Sequences for putative binding sites and their positions were indicated. (B) HEK293T cells were transfected with HRE-luciferase reporter plasmids (P1, P2 or P3) under normoxia or hypoxia. After 48 h, luciferase activity was measured. Transfection efficiency was normalized against Renilla luciferase expression. (C) HEK293T cells were transfected with HRE-luciferase reporter plasmids (P4 or P5) under normoxia or hypoxia. After 48 h, luciferase activity was measured. Transfection efficiency was normalized against Renilla luciferase expression. (D) Schematic diagrams of the regulating sequences with the putative HREs of PIM2. Mut: CG was changed into AA. HEK293T cells were transfected with the HRE (+355mu), HRE (+458mu), HRE (+635mu) or HRE (+789mu)-luciferase reporter plasmids under normoxia or hypoxia. After 48h, luciferase activity was measured. Transfection efficiency was normalized against Renilla luciferase expression. (E and F) Anti-IgG, anti-HIF-1α (E) or anti-HIF-2α (F) antibodies were used in the chromatin immunoprecipitation (ChIP) assays using HEK293T cells which were treated with hypoxia for 12 h. All data represent the means ± SEM of three independent experiments, *p<0.05, **p<0.01.