ELR510444 decreases HIF-1α and HIF-2α expression.

<p>(A) ELR510444 (chemical structure on left) inhibits HIF-1α activity. RCC4 cells were treated with the indicated concentrations of ELR510444 for 16 h. Cells were harvested, nuclei isolated and lysed, and HIF-1α activity was measured using a commercial activity kit according to the manufacturer's protocol. Mean ± SD, <i>n</i> = 3. (B) ELR510444 decreases HIF-1α and HIF-2α protein expression. RCC4 cells were treated with ELR510444 for 24 h and protein levels were measured by immunoblotting. (C) Quantitative real-time PCR analysis of <i>HIF-1α</i> and HIF-2α (<i>EPAS1</i>) levels. RCC cells were treated with 10 nM ELR510444 for 24 h and gene expression was measured by qRT-PCR. Mean ± SD, <i>n</i> = 3. *Indicates a significant difference from controls, p<0.05. Note that HIF-1α is not present 786-O and A498 cells. (D) ELR510444 decreases VEGF secretion. RCC cells were treated with 10 nM ELR510444 for 16 h. Media was harvested and VEGF levels were measured by ELISA. Mean ± SD, <i>n</i> = 3. *Represents a significant difference compared to controls, p<0.05.