To validate the biological roles of 14-3-3γ in regulating PDK1/AKT signaling, we deplete endogenous 14-3-3γ in cells and reveal the potent role of 14-3-3γ in inhibiting AKT kinase by manipulating PDK1 localization and dimerization, thus promoting cancer cell malignant phenotypes. Here, YWHAG is linked to cancer.