Background: The unilateral prone arm lift (UPAL) is commonly used to exercise the lower trapezius muscle. However, overactivation of the upper trapezius can induce pain during UPAL exercises in subjects with upper trapezius tenderness. Objects: The purpose of this study was to investigate the effects of position of ipsilateral neck rotation (INR) on the inhibition of upper trapezius muscle activity and the facilitation of the lower trapezius muscle when performing UPAL exercises. Methods: In total, 19 subjects with upper trapezius tenderness were recruited for the study. Electromyographic (EMG) activity was measured in the upper, middle, and lower trapezius muscles during UPAL with and without INR position. Wilcoxon signed-rank test was used to compare EMG activity in the trapezius muscles and the muscle ratios. Results: EMG activity in the upper trapezius muscles was decreased significantly in the INR condition compared to without the position with INR during UPAL exercises (p<.05). EMG activity in the middle and lower trapezius was not significantly different between the with and without INR conditions (p>.05). However, the ratio of lower to upper trapezius activation showed a significant increase in the INR condition compared to the without INR condition (p<.05), indicating greater lower trapezius activation relative to the upper trapezius in the INR position than in the without INR position. Conclusions: The EMG results obtained in this study suggest that the position with INR reduced overactivation in the upper trapezius and improved muscle imbalance during lower trapezius exercises in individuals with upper trapezius tenderness.