The regulation of cell-cell adhesion is important for the processes of tissue formation and morphogenesis. Here we report that loss of 14-3-3γ leads to a decrease in cell-cell adhesion and a defect in the transport of plakoglobin (PG) and other desmosomal proteins to the cell border in HCT116 cells and in the mouse testis. 14-3-3γ binds to PG in a PKCμ dependent fashion resulting in microtubule dependent transport of PG to the border. Transport of PG to the border is dependent on the KIF5B/KLC1 complex. Knockdown of KIF5B in HCT116 cells or in the mouse testis, results in a phenotype similar to that observed with 14-3-3γ knockdown. Our results suggest that loss of 14-3-3γ leads to decreased desmosome formation and a decrease in cell-cell adhesion in vitro and in vivo in the mouse testis leading to defects in testis organization and spermatogenesis.