The purpose of this study was to investigate the effect of pre-exhaustion exercise on lower-extremity muscle activation during a leg press exercise. Pre-exhaustion exercise, a technique frequently used by weight trainers, involves combining a single-joint exercise immediately followed by a related multijoint exercise (e.g., a knee extension exercise followed by a leg press exercise). Seventeen healthy male subjects performed 1 set of a leg press exercise with and without pre-exhaustion exercise, which consisted of 1 set of a knee extension exercise. Both exercises were performed at a load of 10 repetitions maximum (10 RM). Electromyography (EMG) was recorded from the rectus femoris, vastus lateralis, and gluteus maximus muscles simultaneously during the leg press exercise. The number of repetitions of the leg press exercise performed by subjects with and without pre-exhaustion exercise was also documented. The activation of the rectus femoris and the vastus lateralis muscles during the leg press exercise was significantly less when subjects were pre-exhausted (p < 0.05). No significant EMG change was observed for the gluteus maximus muscle. When in a pre-exhausted state, subjects performed significantly (p < 0.001) less repetitions of the leg press exercise. Our findings do not support the popular belief of weight trainers that performing pre-exhaustion exercise is more effective in order to enhance muscle activity compared with regular weight training. Conversely, pre-exhaustion exercise may have disadvantageous effects on performance, such as decreased muscle activity and reduction in strength, during multijoint exercise.