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Reports on the amount of unloading maximizing walking ability in patients with stroke are limited. The effect of body weight-supported treadmill training (BWSTT) in patients with stroke has not been clarified. We aimed to investigate the effects of unloading rate during BWSTT on the gait of patients with stroke and the relationship between BWSTT and walking ability on flat ground. [Participants and Methods] We performed BWSTT in 17 patients at three unloading rates: 0%, 20%, and 40%. Then, we examined the walking speed and rate, number of steps, single-leg support time ratio, and root mean square before and after unloading. Furthermore, we examined the relationship between walking ability on flat ground and immediate effects of BWSTT at each unloading rate. [Results] We observed no significant improvement under all conditions. However, walking ability improved at unloading rates of 20% and 40%, with poor temporal symmetry while walking on flat ground. [Conclusion] Our results revealed that BWSTT has diverse effects depending on the unloading rate and the ability to walk on flat ground. In particular, it tends to be highly effective for those who have poor sway and symmetry, which may serve as an index for prescribing BWSTT.