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Figure 2


Fig. 2. Abl regulates filopodium formation during attachment through the interaction of PxxP motifs with Crk and Nck family proteins. (A-C) NIH3T3 cells were infected with control retrovirus (derived from pSUPER vector) or ones carrying shRNA sequence targeting CrkI, CrkII and CrkL, or Nck1 or Nck2. These knockdown cells were superinfected with control retrovirus (derived from MIGR-1 vector) or virus expressing shRNA-resistant (human) CrkII or Nck2. (D-F) NIH3T3 cells overexpressing Abl, Nck2 or CrkII were treated with or without STI-571. (A,D) Protein expression was examined by western blotting. (B,F) Serum-starved cells were plated on coverslips coated with 10 µg/ml fibronectin, fixed at 20 minutes after plating, and stained for F-actin and DNA. Scale bars: 20 µm. (C,E) The number of filopodia was counted on fixed cells. Graph represents mean ± s.e.m. number of filopodia per cell or percentage of cells showing filopodia. {ddagger}P<0.05; P=0.143.