Figure 6

PD 81,723 inhibits angiogenic potential, migration, and cell proliferation. (A) PD 81,723 reduced endothelial branching and outgrowth in a matrigel assay. Representative images of HUVECs in matrigel treated with 0.05% DMSO and 50 µM PD 81,723 at 9 h after seeding with a graphical representation of the results from the assay. The error bars represent the standard deviation of the means from the six wells for each condition. The total capillary tube length was significantly reduced in the presence of PD 81,723 (*** = P < 0.001). Scale bars are 100 µm. (B) PD 81,723 inhibits wound closure in HUVECs. The cells were treated with 0.05% DMSO or 50 µM PD 81,723 in either complete endothelial growth media or endothelial growth media without VEGF-A. Five sites near the center of each gap were imaged every 30 min for 24 h. The wounds treated with PD 81,723 in complete media and DMSO in media without VEGF-A took significantly longer to close (* = P < 0.05) than the wounds treated with DMSO in complete media. The wounds treated with PD 81,723 in VEGF-A deficient media did not close within a 24 h period. The error bars represent the standard deviation of the means from three wells. (C) PD 81,723 inhibits proliferation in HUVECs. A bromodeoxyuridine (BrdU) incorporation cell proliferation kit was used to look at cell proliferation. The cells were treated with 0.05% DMSO or 50 µM PD 81,723 in either complete endothelial growth media or endothelial growth media without VEGF-A, with five wells per condition in a 96-well plate. There was no notable difference between DMSO groups, while PD 81,723 significantly (** = P < 0.01 vs. complete media control) reduced cell proliferation in both cases. This experiment was repeated in triplicate. The error bars represent the standard deviation of the means from the three experiments.