When a high power output is required in cycling, a spontaneous transition by the cyclist from a seated to a standing position generally occurs. In this study, by varying the cadence and cyclist bodyweight, we tested whether the transition is better explained by the greater power economy of a standing position or by the emergence of mechanical constraints that force cyclists to stand. Ten males participated in five experimental sessions corresponding to different bodyweights (80%, 100%, or 120%) and cadences (50 RPM, 70 RPM, or 90 RPM). In each session, we first determined the seat-to-stand transition power (SSTP) in an incremental test. The participants then cycled at 20%, 40%, 60%, 80%, 100%, or 120% of the SSTP in the seated and standing positions, for which we recorded the saddle forces and electromyogram (EMG) signals of eight lower limb muscles. We estimated the cycling cost using an EMG cost function (ECF) and the minimal saddle forces in the seated position as an indicator of the mechanical constraints. Our results show the SSTP to vary with respect to both cadence and bodyweight. The ECF was lower in the standing position above the SSTP value (i.e., at 120%) in all experimental sessions. The minimal saddle forces varied significantly with respect to both cadence and bodyweight. These results suggest that optimization of the muscular cost function, rather than mechanical constraints, explain the seat-to-stand transition in cycling.