Inhibition of PMA-induced endothelial cell activation and adhesion by over-expression of domain negative IκBα protein

Figure 1 Determining NF-κB activity in ECV304WT and ECV304MT cells. A: IκBα level were detected after treated with PMA (50 nmol/L) for 12 h; B: EMSA was performed to determine the activity of NF-κB after treated with PMA (50 nmol/L) for 12 h. Band of NF-κB was marked.

Figure 2 Down-regulation of PMA-induced adhesion molecules expression in ECV cells transfected with pCMV-IκBαM compared with pCMV-IκBα. RT-PCR analysis was performed with primers specific for ICAM-1 (A) and VCAM-1 (B). C: The expression of ICAM-1, VCAM-1 and P-selectin were detected using specific antibodies by flow cytometry. Results are representative of at least three independent experiments.

Figure 3 Adhesion of human Jurkat T-cells to PMA-activated endothelial (ECV) cells is inhibited by transfection with pCMV-IκBαM compared with pCMV-IκBα. ECV cells were activated with PMA (50 nmol/L) for 12 h. Cells were washed thrice with PBS and then co-cultured with Jurkat T-cells for 2 h. Then the cells were washed with PBS thrice. Jurkat cell adhesion was determined by visual counting under a phase-contrast microscope. P<0.05 when compared ECVMT with ECVWT.