A single bout of lower-body aerobic exercise may negatively affect a subsequent lower-body resistance exercise workout. However, less is known regarding the effects of a lower-body aerobic workout on muscle activation and performance during a subsequent upper-body resistance exercise workout. Therefore, the purpose of this study was to compare muscle activation and performance during lower- and upper-body resistance exercise workouts after a single bout of lower-body aerobic exercise on an elliptical machine. Fourteen men (mean age = 24.1 ± 2.3 years, height = 180.8 ± 6.9 cm, body mass = 91.9 ± 16.4 kg) completed 4 trials in random order. Two trials consisted of 30 minutes on the elliptical machine, using the lower body only, at 70% of age-predicted maximum heart rate before either a back squat or bench press workout, consisting of 3 sets to failure performed at 75% 1 repetition maximum. The other 2 trials consisted of only the back squat or bench press resistance workouts. To quantify muscle activation, bipolar surface electromyography electrodes were placed on the rectus femoris and vastus lateralis or pectoralis major. Acute lower-body aerobic exercise on an elliptical machine significantly reduced the number of repetitions completed for the back squat but not the bench press exercise. There was no significant difference in muscle activation between the elliptical and no elliptical conditions. However, for both exercises and conditions, muscle activation increased significantly between the first and final repetitions for the first 2 sets but not for the third set. These results suggest that to optimize the quality of a lower-body resistance-training workout, the workout should not be preceded by lower-body aerobic exercise.