Stair locomotion is an important but challenging functional activity for people with lower limb pathology. This study aimed to investigate the bilateral changes in force-bearing on lower limbs during stair locomotion in patients with unilateral ACL deficiency. The ground reaction forces (GRF) were collected from three force platforms: One at ground level in front of a 5-step stair and two on the first two steps respectively. Parameters in vertical and anterior-posterior GRF were extracted and compared between the ACL-deficient (ACLD) and control groups. The ACLD group showed significantly slower stepping cadences in both stair ascent and stepping down to the ground (p < 0.05). The vertical GRF in the ACLD group demonstrated smaller peak forces but larger minimum forces between the two peaks than those in the control group during both stair ascent and descent. Significantly reduced anterior propulsive forces and push-off rates in the late stance were also found in both limbs of the ACLD group (p < 0.05). The slower cadences and reduced force-bearing on the affected limb suggested a protective strategy was adopted. However, the anterior loading parameters in the early stance on the unaffected limb demonstrated different adaptations with significantly larger magnitudes during stair ascent but reduced magnitudes during stair descent (p < 0.05). Similar results were also found in the weight- transferring strategies between legs in consecutive steps with a significantly larger percentage of lift-up forces but a smaller percentage of impact forces on the leading unaffected limb. The results of this study indicated a cautious force-bearing strategy and bilateral adaptation were apparent in the patients with unilateral ACL deficiency. This information may provide a safety guideline for the patients and be helpful for a better use of the stair tasks as part of a rehabilitation program.