Figure 5.

Overexpression of pVHL does not change HIF activity. A) N12 cells were cotransfected with a luciferase reporter construct driven by phosphoglyceric kinase promoter containing HREs and Renilla luciferase reporter plasmid. Then cells were infected with Ad-Null or Ad-VHL-HA and incubated for 24 h. Another group of cells was exposed to hypoxia (1.5% O₂) after cotransfection of HREs and Renilla reporter plasmids. Dual luciferase assay (Promega) was carried out following the manufacturer's protocol. HRE luciferase activity was normalized to Renilla luciferase activity. Relative luciferase activity of uninfected group was set to 1; 3 independent experiments. B) N12 human primary lung fibroblasts were infected with pAd-Null or pAd-VHL-HA for 2 d, and mRNA expression levels of VEGF were determined by qRT-PCR; RPL19 gene was used as internal control. Relative mRNA level of VEGF of uninfected group was set to 100; 4 independent experiments. Graphs represent means + se. **P < 0.01.