Fig. 1.

EphrinB2 and EphB4 increase VEGF mRNA levels in human hepatic stellate cells (HSC). HSC were treated with EphrinB2 chimera (2 μg/ml), EphB4 chimera (2 μg/ml), and/or respective small interfering RNA (siRNA) for 3, 12, and 48 h, respectively, and the total RNA was extracted for VEGF mRNA estimation by quantitative RT-PCR analysis. A: there was a significant increase in VEGF mRNA levels in both EphrinB2- and EphB4-treated groups compared with their respective time controls at 12 and 48 h (n = 3 separate experiments, each in triplicate, *P ≤ 0.05). B: cyclohexamide (10 μg/ml) blocked EphB4 Fc-induced VEGF A mRNA production (n = 3 separate experiments, each in triplicate, *P ≤ 0.05). C: basal VEGF mRNA levels were attenuated by incubating HSC with EphrinB2 or EphB4 siRNA (n = 3 separate experiment, each in triplicate, *P ≤ 0.05). Stimulation of siRNA-transfected cells with EphrinB2 and EphB4 chimeras reversed the siRNA-mediated inhibitory effect on VEGF mRNA levels.