Cyclists with unilateral transtibial amputation (CTA) provide a unique model to study integration of the neuromuscular and bicycle systems while having the option to modify this integration via the properties of the prosthesis. This study included eight CTA and nine intact cyclists. The cyclists pedaled on a stationary bicycle with instrumented force pedals. The CTA group pedaled with a stiff or flexible prosthetic foot during a simulated time trial and a low difficulty condition. During the time trial condition, pedaling with the flexible foot resulted in force and work asymmetries of 11.4% and 30.5%, the stiff foot displayed 11.1% and 21.7%, and the intact group displayed 4.3% and 4.2%, respectively. Similar trends were shown in the low difficulty condition. These data suggest foot stiffness has an effect on cycling symmetry in amputees.