Click on image to view larger version.
Figure 1. Eap interferes with angiogenic response of EC primary cultures. (A) mRNA levels of DAF and TF were determined by real-time polymerase chain reaction (PCR) in HUVEC stimulated with either 100 ng/ml VEGF or bFGF in the absence (open bars) or presence (solid bars) of 20 µg/ml Eap. Results are shown as fold increase over basal expression (means±SD) of three independent experiments and shown as *P < 0.05; **P < 0.01; ***P < 0.001. (B) HUVEC were grown to subconfluence in 24-well plates, serum-starved overnight (0.5% FCS), and then stimulated with either VEGF (25 ng/ml), bFGF (25 ng/ml), or medium alone (0.1% FCS and 0.5% BSA) for 20 h in the absence (solid bars) or presence (open bars) of 20 µg/ml Eap. Cells were pulse-labeled with tritium-thymidine for another 4 h and analyzed for thymidine incorporation. Data represent mean counts per minute ± SD with n = 5; *P < 0.05; **P < 0.01; ***P < 0.001. (C) Eap inhibits capillary sprout formation. Capillary-like sprout formation of BREC was analyzed in the absence (open bars) or presence (solid bars) of 2 ng/ml bFGF and with the indicated concentrations of isolated Eap. Data represent the mean ± SD of 30 beads analyzed in triplicate each.
