FIGURE 1.

Co-localization and direct interaction between Gas7 and N-WASP in vitro. The microtubule organization (A), actin reorganization, and co-localization of Gas7 and N-WASP (B) in transfected neuroblastoma cells is shown. Neuro-2a cells were transfected only with Gas7/Myc or co-transfected with HA/N-WASP to examine the distribution of microtubule, actin, Gas7, and N-WASP. Gas7, N-WASP, and actin filaments are co-localized at the submembrane region as well as in protrusions (arrows). Scale bar, 5 μm (2-μm inset). C, interaction of Gas7 and N-WASP in cells is shown. Extracts from COS-7 cells co-transfected with Gas7/Myc and HA/N-WASP were analyzed by co-immunoprecipitation (IP). N-WASP was only detected in association with Gas7. Antibodies against Gas7 were raised in rabbits, and preimmune serum (PIS) was used as a negative control. D, direct interaction between Gas7 and N-WASP in vitro is shown. Recombinant GST-Gas7 and His-NWASP were used in a GST pulldown assay to examine the interaction between Gas7 and N-WASP. His-N-WASP was directly pulled down by immobilized Gas7 but not by GST alone (negative control).