This study aimed to evaluate the effect of arm posture on activation of the anterior and posterior regions of supraspinatus and the superior and middle regions of infraspinatus during resisted isometric arm elevations.Thirty-one healthy participants performed eighteen isometric resistance exertions against a robotic arm in three elevation planes (flexion, scaption, abduction) and three elevation angles (30°, 90°, 150°) in maximal and sub-maximal resistance conditions. EMG data were obtained using fine wire electrodes. The mean activation of each region and the activation ratios were compared across postures using ANOVAs.Supraspinatus anterior was significantly more active during abduction and scaption, and in higher elevation angles, while the posterior region showed similar activation levels across postures. Infraspinatus regions were more active during flexion with more relative activation of the infraspinatus superior at 90° flexion.The results suggest that regional activation of supraspinatus and infraspinatus should be considered for assessment and rehabilitation purposes. In any clinical condition that less stress on the supraspinatus anterior is desired, isometric training in flexion or in lower elevation levels may strengthen the supraspinatus posterior while causing lower stress in the anterior region.Beside external rotation exertions, resisted flexion tests may be useful for evaluation of infraspinatus regions.