Neutron reflectometry measurements show that lamellar structures composed of thin alternating water-rich and Nafion-rich layers exist at the interface between SiO2 and the hydrated Nafion film. Lamellae thickness and number of layers increase with humidity. Some lamellae remain in the film after dehydration. Multilayer lamellae are not observed for Nafion on Au or Pt surfaces. Instead, a thin partially hydrated single interfacial layer occurs and decreases in thickness to a few angstroms as humidity is reduced to zero. The absorption isotherm of the rest of the Nafion film is similar to that of bulk Nafion for all three surfaces investigated. The observed interfacial structures have implications for the performance, reliability. and improvements of fuel cell proton exchange membranes and membrane electrode assemblies.