The musculoskeletal models have been improved to estimate accurate knee compression force (KCF) and have been used to reveal the causal relationship between KCF and muscle weakness. Previous studies have explored how muscle weakness influences the KCF during gait; however, the influence of muscle weakness is possibly larger during activities that require deeper knee flexion (e.g., stair ambulation) than other activities (e.g., gait) because of the small knee contact area of articular surfaces. To explore how muscle weakness influences the KCF during stair ambulation. Ten young adults performed stair ascent and descent tasks at a comfortable speed. Based on a previous study, we created muscle weakness models of rectus femoris (RF), vastus muscles (VAS), gluteus medius (Gmed), and gluteus maximus (Gmax), and the medial and lateral KCF (KCFmed and KCFlat) during stair ambulation were calculated. Similar to the gait, the Gmed weakness increased KCFmed and decreased KCFlat during stair ascent and descent. Whereas, unlike the gait, the Gmax weakness increased KCFmed during stair ascent and the VAS weakness decreased KCFmed and KCFlat during stair ascent and descent. Moreover, the percentage changes in KCF were similar (or large) during stair ambulation compared with those during gait. Considering the KCF alterations caused by each muscle weakness, the weaknesses in Gmax and Gmed might lead to cartilage loss and pain in the knee, and the VAS weakness might lead to low stability of the knee. The symptom during stair ambulation might help precisely identify the muscle requiring rehabilitation. •Musculoskeletal models is useful to explore how muscle weakness affects knee compression force.•We explored which muscle weakness influences knee compression force during stair ambulation.•Gluteus medius and maximus weaknesses induced high medial and low lateral knee compression force.•Vastus weakness induced low medial and lateral knee compression force.