A number of factors, including protein kinases, Rho GTPases and actin and microtubule cytoskeletons play a crucial role in cell migration and spreading. We have recently shown that ectopic expression of FAM110C can alter cellular morphology by mechanisms yet to be determined. In this study, a FAM110C antiserum has been developed and used to study endogenously expressed FAM110C. Our data show that FAM110C is expressed by different cell lines and it can be detected throughout the cell. Interestingly, depletion of FAM110C by short interfering RNA reduced integrin-mediated filopodia formation, hepatocyte growth factor-induced migration, and phosphorylation of the Akt1 kinase in the epithelial cell line HepG2. Furthermore, co-immunoprecipitation and co-localization studies show that both ectopically and endogenously expressed FAM110C interact, or is part of a protein complex, with the Akt1 kinase. This interaction is transient and follows the activation of Akt1. In addition, we show that alpha-tubulin co-precipitates with FAM110C which further supports an interaction with the microtubule cytoskeleton. Collectively, these findings suggest a new function for FAM110C in the regulation of cell spreading, migration and filopodia induction.