Human navigation heavily relies on visual information. Although many previous studies have investigated how navigational information is inferred from visual features of scenes, little is understood about the impact of navigational experience on visual scene representation. In this study, we examined how navigational experience influences both the behavioral and neural responses to a visual scene. During training, participants navigated in the virtual reality (VR) environments which we manipulated navigational experience while holding the visual properties of scenes constant. Half of the environments allowed free navigation (navigable), while the other half featured an 'invisible wall' preventing the participants to continue forward even though the scene was visually navigable (non-navigable). During testing, participants viewed scene images from the VR environment while completing either a behavioral perceptual identification task (Experimentl) or an fMRI scan (Experiment2). Behaviorally, we found that participants judged a scene pair to be significantly more visually different if their prior navigational experience varied, even after accounting for visual similarities between the scene pairs. Neurally, multi-voxel pattern of the parahippocampal place area (PPA) distinguished visual scenes based on prior navigational experience alone. These results suggest that the human visual scene cortex represents information about navigability obtained through prior experience, beyond those computable from the visual properties of the scene. Taken together, these results suggest that scene representation is modulated by prior navigational experience to help us construct a functionally meaningful visual environment.