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Figure 6


Fig. 6. Augmented adhesion of EphB4-overexpressing monocytic cells to ephrin-B2-overexpressing ECs. (A) Western blot analysis of mock-transduced mouse J774 cells (J774), EphB4-GFP-overexpressing J774 cells (J774 EphB4) and {Delta}C-EphB4–GFP-overexpressing J774 cells (J774 {Delta}C-EphB4). All three cell populations expressed endogenous EphB4 as reflected by a faint, but detectable band at 130 kDa (middle arrow). Full-length EphB4-GFP (>130 kDa) and truncated {Delta}C-EphB4–GFP (<130 kDa) are marked accordingly. Actin served as loading control. (B) Adhesion of mock-transduced, and EphB4- and {Delta}C-EphB4-overexpressing J774 cells to ephrin-B2-overexpressing and mock-transduced HUVECs. Cells were incubated for 30 minutes on a rocking platform. Endogenous EphB4 expression facilitated the preferential adhesion to ephrin-B2-overexpressing HUVECs compared with mock-transduced HUVECs, confirming the findings obtained with J774 cells (Fig. 3). Overexpression of full-length EphB4 but not {Delta}C-EphB4 in J774 cells further augmented the preferential adhesion of J774 cells to ephrin-B2-expressing HUVECs. Values are expressed as the mean ± s.d. of one out of three independent experiments with similar results (**P<0.01).