FIGURE 11.

CDDO-Im disrupts PKCζ localization at the leading edge of polarized cells. Rat2 fibroblast monolayers were scratched and allowed to grow into the wound for 6 h in order to polarize before being incubated in the absence (Control; A) or presence of 1 μm CDDO-Im (B) or 10 μm nocodazole (C) for an additional 2 h. Cells were then fixed, permeabilized, and immunostained for endogenous Rac1 (green) or PKCζ (red), using anti-Rac1 and anti-PKCζ antibodies, respectively. A representative area of interest (white box) from each condition was enlarged and shown (inset). The green and red arrows indicate Rac1 and PKCζ at the leading edge of migrating cells, respectively. The co-localization of Rac1 with PKCζ is indicated by yellow arrowheads. The blue arrows indicate the direction of movement. Representative images from four experiments are shown. Bar, 10 μm.