Fig. 4. Actin cytoskeleton regulates localization of CD44H and MT1-MMP. CHO-K1 cells were transiently transfected with expression plasmids for CD44H-F (A), MT1-F (B), MT1-F/dPEX (C) and CD44H/dCP + MT1-F (D and E). Transfected cells were seeded on fibronectin-coated glass coverslips for 1 h and treated with anti-FLAG antibody (A–C and E) or anti-CD44 antibody (D) for 30 min at 37°C. After excess antibody had been washed off with PBS, the cells were treated with CyD (1 µg/ml) for 30 min at 37°C. They were then fixed, and the localization of the bound antibody and actin cytoskeleton was visualized with Cy3-conjugated anti-mouse IgG (red) and Alexa488-conjugated phalloidin (A–C and E) or Alexa488-conjugated anti-rabbit IgG and Alexa594-conjugated phalloidin (D), respectively. Scale bar, 10 µm.