FIGURE 8.

Subcellular targeting of biotinylated CDDO. Rat2 fibroblasts were grown to 100% confluence and scratched to create a wound. After incubation for 6 h to allow cell polarization and migration, cells were fixed, permeabilized, and incubated with monoclonal anti-Rac1 antibodies (Rac1; green) and biotin (A), CDDO (B), or CDDO-biotin (C), followed by Cy2-labeled anti-mouse antibody and Cy3-labeled streptavidin. The co-localization of Rac1 (green) with CDDO-biotin (red) at the leading edge of migrating cells is demonstrated in the inset (yellow arrowheads). The blue arrow indicates the direction of cell movement, and DIC microscopy was included to visualize the leading edge of migrating cells. Representative images from four experiments are shown. Bar, 10 μm.