Thrombin-induced ICAM-1 upregulation was dependent on EphA2. HUVECs cells were transfected with either a control siRNA or one of the two EphA2-specific siRNA. Confluent HUVECs were stimulated with 1U/ml thrombin (T) for 6 hours. (A) Total lysates were used for western blots. Effective expression knockdown by both EphA2-specific siRNAs, thrombin-induced ICAM-1 expression, and protein loading were examined by western blot analyses using antibodies specific to EphA2, ICAM-1, and α-actinin, respectively. Representative data from 3 experiments are shown. (B) Densitometry analysis of ICAM-1 upregulation by thrombin in the presence or absence of EphA2. Western blots were scanned and analyzed by using the program ImageJ. Results were reported as fold-increase in ICAM-1 expression relative to unstimulated controls. Data are mean ± s.d. of three experiments; * p<0.002.