FIG.8.

Coexpression of tyrosine-phosphorylated Dab1RFP and Nckβ in Rat-2 cells leads to the disruption of the actin cytoskeleton. The actin cytoskeleton, detected with fluorescently labeled phalloidin (white; B, D, F, H, J, and L) observed on the lower plane of Rat-2 fibroblasts transfected with HA-Nckβ (green; A and B), or Dab1RFP, a tyrosine-phosphorylated form of Dab1 (red; C and D), shows the typical mesh pattern. Combined expression of HA-Nckβ and Dab1RFP (E and F) leads to disruption of the actin mesh and clumping of actin filaments. Expression of unphosphorylated Dab1-wt with HA-Nckβ (G and H), or Dab1RFP-5F with HA-Nckβ (I and J) did not disrupt the actin cytoskeleton, nor did expression of Dab1RFP with the SH2 domain mutant of Nckβ (K and L). All images show the plane of the cell closest to the cover glass. Bar, 20 μm.