Fig. 1. Immunolocalization of N-WASP at different stages of the cell cycle. (A) HeLa cell lysate was immunoblotted with purified anti-N-WASP antibody. Flag-tagged full-length N-WASP expressing plasmid (Flag-NW) and empty vector (Flag-vector) alone were also transfected into HeLa cells. Whole cell lysates for these cells were immunoblotted with anti-Flag or anti-N-WASP antibody. (B) HeLa cells were fixed with-20℃ methanol at different stages of cell cycle progression and stained for α-tubulin, TOPRO3, and N-WASP. In prometaphase and metaphase, N-WASP was seen as a pattern of dots reminiscent of kinetochores. In interphase and rophase, N-WASP is concentrated in the nucleus. In telophase and cytokinesis, intense signals of N-WASP were observed at midbody.