Figure 6.

Pharmacological activation of HIF-1α downregulates Cav-1 levels. (A) Immunofluorescence. to pharmacologically activate HIF-1α, co-cultures of hTERT-fibroblasts and MCF7 cells were treated with PHD inhibitors, such as DMOG, 2,4 DPD and 1,4 DPCA. Cells were then fixed and immuno-stained with anti-Cav-1 (red) and anti-K8-18 (green) antibodies. Nuclei were stained with DAPI (blue). Cav-1 staining (red channel only) is shown on the left to better appreciate the decreased Cav-1 levels, after treatment with the PHD inhibitors. Importantly, images were acquired using identical exposure settings. Original magnification, 40x. (B) Western blot. Homotypic cultures of hTERT-fibroblasts were treated with the PHD inhibitor DMOG (500 µM) or vehicle control (DMSO) for 24 hours. Cell lysates were analyzed by western blot analysis using anti-Cav-1 antibodies. Note that Cav-1 levels are greatly decreased upon treatment with the HIF-1α inducer DMOG. β-actin was used as a control for equal protein loading.