This descriptive study aimed to compare the magnetic resonance appearance of the distal interphalangeal joint articular cartilage between standing weight-bearing and non-weight-bearing conditions. Ten forefeet of live horses were scanned in a standing low-field magnetic resonance system (0.27 T). After euthanasia for reasons unrelated to the study, the non-weight-bearing isolated feet were scanned in a vertical positioning reproducing limb orientation in live horses. The same acquisition settings as during the weight-bearing examination were used. Thickness and cross-sectional area of the distal interphalangeal articular cartilage and joint space were measured on tridimensional T1-weighted gradient echo high resolution frontal and sagittal images at predetermined landmarks in both conditions and were compared using a linear mixed-effects model. Frontal images were randomized and submitted to 9 blinded readers with 3 different experience levels for identification of weight-bearing versus non-weight-bearing acquisitions based on cartilage appearance. Weight-bearing limbs had significantly thinner distal interphalangeal cartilage (p = 0.0001) than non-weight-bearing limbs. This change was greater in the distal phalanx cartilage than that of the middle phalanx. Blinded readers correctly identified 83% (range 65 to 95%) of the images as weight-bearing or non-weight-bearing acquisitions, with significantly different results observed among the different readers (p < 0.001) and groups (p < 0.001). These results indicate that distal interphalangeal articular cartilage and particularly cartilage of the distal phalanx thins when weight-bearing compared to the non-weight-bearing standing postmortem conditions and suggest that cartilage abnormalities may be more difficult to identify on weight-bearing standing magnetic resonance imaging.