ABSTRACT The present work investigates anisotropic fatigue properties of an additively manufactured (AM) medium-entropy alloy (MEA). The fatigue properties of the materials in 0°, 45° and 90° orientations with respect to the build layers were measured. In order to discover the dominant factors for stress level-dependent fatigue anisotropy, crystal plasticity finite element simulations and fatigue crack growth (FCG) experiments were used to evaluate crack initiation and crack propagation behaviours, respectively. The main conclusions are summarised as follows: First, in comparison to grain anisotropy, the difference in fatigue initiation stage is controlled by defect anisotropy. Second, the direct cause of the difference in the FCG rate is considered to be the differences in deflection angle, affected by the incompatibility of the slip planes between adjacent grains. Third, the AM-MEA displays a stress level-dependent fatigue anisotropy. At high-stress level, the fatigue life of the MEA-45° specimen is significantly higher than that of MEA-0° and MEA-90°, while at low-stress levels, the fatigue resistance of MEA-0° is similar to that of MEA-45°. Stress level-dependent fatigue anisotropy is controlled by different ratios of crack initiation and crack propagation. Our work proposes a novel research strategy that qualitatively evaluates fatigue anisotropy of AM materials.