Figure 3
The increase of SLA determinant in HBx-transfected cells and the adhesion of HBx-transfected cells to endothelial cells. (A) The cells were fixed, and then immunostained with SLA and SLX antibodies. The immunostained cells were analyzed using a fluorescence microscope. (B) Chang, Chang-pcDNA and Chang-pcDNA-HBx cells (5 × 105 cells/well) were added on TNF-α-unstimulated or stimulated HUVECs, and were incubated for 20 min at 37°C with rotation at 60 rpm in a shaking incubator. The attached cells were visualized using a microscope. (C) To check the antibody-mediated inhibition of cell adhesion, the TNF-α-stimulated HUVECs were incubated with E-selectin antibody for 4 h at 37°C in a humidified CO2 incubator, and each cell was then added to monolayer-cultured TNF-α-stimulated HUVECs. To block cell adhesion by SLA or SLX-specific antibody, each of cells was incubated with the antibody for 1 h and added to the TNF-α-induced HUVECs.