Figure 9. Role of PKC δ, ε, and ζ isoforms on S1P mediated chemotaxis and lamellipodial localization of Coronin 1B in human lung endothelial cells.

HPAECs grown on slide chambers or 35-mm dishes (∼70% confluence) were infected with empty vector or adenoviral vectors encoding PKC dominant negative (dn) δ, ε, and ζ isoforms (5 MOI) in complete EGM-2 medium for 24 h. (A), Cell lysates (20 µg of protein) were subjected to 10% SDS-PAGE, Western blotting and probed with anti-PKC δ, ε, ζ and actin antibodies. In parallel experiments, the effect of dn PKC δ, ε and ζ isoforms on chemotaxis (B) and lamellipodial localization of coronin 1B and actin (C, D and E) was examined as described in Materials and Methods. Values are mean±SEM of three independent experiments. *, p<0.01 compared cells without S1P; **, p<0.005 compared to cells infected with empty vector and stimulated with S1P.