FIGURE 3.

RhoA is activated by complement in GEC. A, schematic representation of the Raichu1298x RhoA FRET probe. RBD, Rho-binding domain of rhotekin; CFP, cyan fluorescent protein. B, GEC were plated onto glass bottom dishes coated with collagen, transfected with the FRET Raichu RhoA probe. Cells were incubated with complement, and FRET images were captured as described under “Experimental Procedures.” The corrected FRET values of each pixel were converted to pseudocolors. Ten cells per condition from three independent experiments were analyzed, and representative cells are shown. Complement (NS) induced RhoA activation starting at ∼15 min. Control cells (HIS) did not show changes in RhoA activity.