Figure 3
From: Hypoxia and estrogen are functionally equivalent in breast cancer-endothelial cell interdependence
Hypoxia and estrogen induce VEGF in TG1-1 cells. Western blots of TG1-1 cytoplasmic lysates show that hypoxia (A) and estradiol (B) lead to an increase in VEGF. However, treatment of cells with Fulvestrant lead to a reduction in VEGF and treatment with YC-1 restored VEGF to control levels. Conditioned media from TG1-1 cells grown in 6 well plates was harvested and VEGF concentration was determined by ELISA. Corresponding cell pellets were harvested, washed, and analyzed for total protein content. Values shown are expressed as percent differences compared to control TG1-1 conditioned media normalized for the amount of protein in the cell pellet. Similarly to observed findings in the western blot analysis, cells treated with estrogen secreted significantly more VEGF when compared to control, an effect abrogated by YC-1 (C). Similarly cells grown under hypoxic conditions (1% O2) also secreted significantly more VEGF than controls. Data represents two separate experiments performed in duplicate. (*p<.05).