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The aim of this study was to understand and measure the lower limbs muscular activation patterns both in healthy and spinal cord injured (SCI) subjects during robot-assisted locomotor exercise. Electromyographic (EMG) activity of four leg's muscles (rectus and biceps femoris, tibialis anterioris and gastrocnemius) was recorded and analyzed at two different percentages of body weight support, three stepping velocities and three different modalities. SCI subjects were recorded also after four weeks training to evaluate the effectiveness of lower limb robot-assisted rehabilitative treatment. A multi-factor ANOVA on the integrated muscle activity (IEMG) parameters both in healthy and SCI subjects was performed. Higher muscular activities both in healthy subjects and SCI patients were found during the exercises using the "DGO active" modality and higher stepping velocities. A significant increased bilateral muscular activity was observed in each SCI subject after the rehabilitation treatment. The method proposed to analyze EMG data provides a quantitative description of the lower limb muscular recruitment and can contribute to identify the optimal rehabilitation treatment's conditions.