Figure 1
Pin1 knockdown upregulates HIF-1α and facilitates HIF-1-driven gene expression. (a) HEK293 cells were transfected with the indicated siRNAs, incubated in normoxia or hypoxia (1% O2) for 8 h, and subjected to western blotting. HIF-1α blots were quantified using ImageJ and displayed in bar graph (mean ± SD, n = 3). #P < 0.05 compared to the control group; *P < 0.05 by Student’s t-test. (b) Cells were treated as in (a) and HIF1A mRNA expression was measured by RT-qPCR. Data represent mean ± SD (n = 3). #P < 0.05 compared to the control group; *P < 0.05 by Student’s t-test. (c) Cells were transfected with the EPO enhancer (or VEGF promoter)-luciferase plasmid, the CMV-β-galactosidase plasmid, and the indicated siRNAs. After cells were incubated in normoxia or hypoxia for 16 h, luciferase activities were measured, and normalized to β-galactosidase activities. Data represent mean ± SD (n = 3). *P < 0.05 by Student’s t-test. (d) Cells were transfected with the indicated siRNAs, incubated in normoxia or hypoxia for 16 h, and lysed for RNA extraction. BNIP3, CA9, LOX, and PDK1 mRNA expression levels were measured by RT-qPCR. Data represent mean ± SD (n = 3). *P < 0.05 by Student’s t-test.
