Wood is a hygroscopic material. Increasing moisture decreases the macroscopic mechanical properties of wood. Investigations of the mechanical behaviour of the solid wood substance at the cell wall scale may contribute to an enhanced insight into moisture-mechanics relationships. Wood cells walls are composed of several different layers, of which the S2 layer and the middle lamella (ML) are mechanically most important. Their mechanical properties can be assessed by nanoindentation. Wood of two hardwood species and three softwood species is investigated. Mechanical properties of the S2 layer as well as of the ML are determined at different relative humidity (RH), i.e. at different wood moisture contents (MC).