Electrospun nanofiber membranes show high potential in various application fields (e.g., filtration, catalysis, and sensing). Nevertheless, knowledge of the mechanical behavior, and more specifically, the deformation of nanofiber membranes is still limited today which can complicate the appliance of nanofiber membranes in applications where they are mechanically loaded. In this paper, we, therefore, analyzed the mechanical behavior of polymeric nanofiber membranes with different fiber orientations (random and aligned) extensively. Polyamide 6 was used as a representative reference polymer for proof-of-concept. Mechanical tests show that all membranes have a coherent deformation behavior at the macroscale up to the point of fracture. Large variations in stiffness, ultimate strength, and ultimate strain were observed between membranes with different fiber orientations (Random: E-mod: 370 ± 34 MP; UTS: 38.5 ± 6.0 MPa; ε_max: 30.0 ± 2.8%; Parallel aligned: E-mod: 753 ± 11 MPa; UTS: 55.4 ± 0.8 MPa; ε_max: 12.0 ± 0.1%; Perpendicular aligned: E-mod: 24.1 ± 3.7 MPa; UTS:/; ε_max: >40%). This shows the versatility and tunability of the mechanical behavior of these nanofiber membranes. At the microscale, the fibrous structure results in deformation mechanisms that resist failure formation and progression when the membrane is mechanically loaded. This results in a high fracture resistance, even for pre-damaged membranes. Realignment of the fibers along the loading direction causes crack tip blunting, locally reinforcing the membrane.