Figure 1.

In vitro proliferation of A2780 stable clones and binding properties of PlGF2-DE. A. A2780 stable clones were seeded in 96-well plates (2000 cells/ cm²) and cell proliferation was evaluated using the CellTiter Aqueous One Cell Proliferation Assay (Promega) at indicated time. B. Binding of VEGF-A homodimer (2.5 to 10 ng/ml), VEGF-A/PlGF2-DE heterodimer (7.5 to 30 ng/ml) and PlGF2-DE homodimer (81.25 to 325 ng/ml) present in the supernatant of A2780-PlGF2-DE stable clone, to coated Flt-1 (0.5 μg/ml) in ELISA based assay. For both experiments each point was carried out in triplicate and the data are represented as the mean of two experiments ± SEM. (C) Western blot analysis of Flt-1 phosphorylation (anti-P-Flt-1) induced by 20 ng/ ml of PlGFwt isoforms or 100 ng/ml of PlGF-DE isoforms, on starved 293-Flt-1 cells. One hundred μg of the cell protein extracts were analyzed. Normalization was performed with ant-Flt-1 antibody.