These data demonstrate the regulation of the Rac to RhoA GTPase switch is critical for cancer cell invasion, and further support the model prediction that a Sos1-Ras-Raf-MEK-ERK negative feedback loop affects Rac1 and RhoA via the Sos1-Eps8-Abi1 complex to determine the mode of cell migration in 3D microenvironments. Here, EPS8 is linked to cancer.