Ect2 Is Recruited by Caveolae to Activate RhoA
(A) Control (left) and Ect2 knockdown (center) A2780 cells expressing membrane-targeted Raichu-RhoA in 3D CDM; right: average forward rear movement over 5 min of control, Ect2, Vav2, and PLEKHG5 knockdown cells (N>46 cells/condition, 3 repeats).
(B) Ratiometric FRET imaging of control, Ect2, Vav2, and PLEKHG5 knockdown cells; average rear RhoA FRET ratio in control, Ect2, Vav2, and PLEKHG5 knockdown cells (N>32 cells/condition, 3 repeats).
(C) Endogenous Ect2 and cavin-1 in A2780 cells in 3D-CDM; MIPs of whole cell and separately captured zoomed rear region. Right: peak Ect2 staining intensity in rear, nuclear, and front regions (N=17 cells, 3 repeats).
(D) Control (left) and caveolin-1 knockdown cells (right) in CDM stained for F-actin (phalloidin) and Ect2, MIPs shown; Rear peak/front peak Ect2 intensity levels (N>20 cells/condition, 3 repeats).
∗∗∗p < 0.001; ∗∗∗∗p < 0.0001; ns, not significant. See also Figure S5.