FIGURE 12.

CDDO-Im disrupts Par6 and TGFβ receptor localization at the leading edge of migrating cells. Rat2 fibroblast monolayers were scratched and allowed to grow into the wound for 6 h before being incubated in the absence (Control), or presence of 1 μm CDDO-Im for an additional 2 h. Cells were then fixed, permeabilized, and immunostained for endogenous Rac1 (green) and Par6 (red) using anti-Rac1 and anti-Par6 antibodies (A) or for endogenous Rac1 (green) and TβRII (red) using anti-Rac1 and anti-TβRII antibodies, respectively (B). A representative area of interest (white box) from each condition was enlarged and shown (inset). The green arrowheads indicate Rac1, and red arrowheads indicate Par6 or TβRII at the leading edge of migrating cells. The co-localization of Rac1 with Par6 or TβRII is indicated by yellow arrowheads. The blue arrows indicate the direction of movement. Bar, 10 μm. Cells containing Rac1, Par6, or TβRII at the leading edge were quantitated from three experiments carried out as described in A and B ± S.D. and graphed (C).