Forkhead box O (FoxOs) is a forkhead family transcription factor which play important roles in differentiation, metabolism, proliferation and survival. FoxO1 and FoxO3a are the members of FoxOs. In the present study, we investigated the role of FoxOs in the exspression of GAP-43 and its underlying mechanisms. Our results showed that overexpression of FoxO1 and FoxO3a inhibited NGF-induced expression of growth associated protein 43 (GAP43), a neurite formation, regeneration, and plasticity related protein, in PC12 cells, while knocking down the expression of FoxO1/3 had the opposite effect. Bioinformatic studies found that the regulatory region of GAP43 promoter contained multiple FoxO3a binding sites. Dual-luciferase reporter assay with report gene containing GAP43 promoter showed that FoxO3a significantly decreased the transcription activity of GAP43 promoter. These results indicate that GAP43 is a direct downstream target of FoxOs which negatively regulates the expression of GAP43. These results put together demonstrate that FoxOs inhibits NGF-induced expression of GAP43 via the PI3K/Akt and MAPK/ERK pathways.