Background: Biomechanical studies indicate that the tibia shifts medially and has a more valgus orientation in the anterior cruciate ligament (ACL)–deficient knee. However, it is not known whether these differences can be detected on standing radiographs. Purpose: To determine whether medial subluxation and more changes in coronal alignment of the tibia are detectable in both weightbearing radiographs and a cadaveric model simulating quiet standing. Study Design: Case series; Level of evidence, 4, and Descriptive laboratory study. Methods: Radiographic data were available for a cross-section of 74 patients with unilateral ACL tears. Tibial subluxation and coronal limb alignment were measured on hip-to-ankle weightbearing radiographs. Eight cadaveric knees were mounted on a 6 degree of freedom robot. Mediolateral position and varus-valgus alignment of the tibia relative to the femur were measured in response to 300-N axial compression simulating quiet standing at 5° and 15° of flexion with the ACL intact and sectioned. Results: Across all 74 patients included in the clinical study, the ACL-injured knee experienced 1.6 ± 2.3 mm (mean ± SD) of medial tibial subluxation compared with the contralateral uninjured knee (P < .001). The 24 patients with isolated ACL rupture exhibited 2.0 ± 1.8 mm of medial subluxation (P < .001). The mean coronal alignment of all 74 patients in the study was 0.7° ± 2.8° varus in the injured limb and 1.3° ± 2.6° varus in the uninjured contralateral limb (P = .0187). In the cadaveric model, the tibia translated 0.4 ± 0.5 mm more medially after sectioning of the ACL at 15° of flexion (P = .0485); however, no changes in coronal alignment were detected. Conclusion: The tibia shifts medially and is less varus in the ACL-deficient knee on standing radiographs. The medial tibial shift is reproduced in an axially loaded cadaveric model. Clinical Relevance: Medial tibiofemoral subluxation seen on frontal plane standing radiograph is an underappreciated sequela of isolated ACL rupture. The ability of ACL reconstruction to restore this aspect of ACL injury is not well understood and should be investigated further. Cadaveric models may be used to directly measure the mechanical effect of subtle changes in mediolateral position on articular contact stress as an indicator of the importance of this finding.