To examine the effect of ankle position (i.e. gastrocnemius muscle length) on training outcomes during leg curl exercise, we recruited untrained and trained healthy adults to participate in two separate experiments. In Experiment 1, we studied the acute influence of ankle position on knee flexor myoelectric (EMG) activity during leg curl exercise in a group of trained and a separate group of untrained adults. In Experiment 2, we studied the effects of ankle position on knee flexors muscle thickness and torque across a 10-week training protocol in trained adults. We hypothesized that leg curl exercise with the ankle in a plantarflexed position would enhance EMG activity, muscular strength, and hamstrings muscle thickness. We randomized the legs within a person to perform leg curl exercise with one in a plantarflexed position and the other in a dorsiflexed position. Experiment 1 revealed no significant differences between ankle positions in the EMG activity of hamstring muscle in either group (all p > 0.05). Experiment 2 revealed a significant pre-to post-intervention increase in biceps femoris long head (BFLH) muscle thickness (p = 0.026) and isometric torque (p = 0.03), but there were no significant effects of the ankle position (p = 0.596) or interaction between ankle position and timepoint for these variables (p = 0.420). In sum, the ankle position did not have acute effects on hamstrings EMG activity, nor did it affect strength and hypertrophy adaptations after 10-weeks of leg curl exercise training. Interestingly, however, the limb which performed leg curl exercise in a dorsiflexed position performed a higher total training volume.