Background: Previous studies have shown that a cerebrovascular accident disrupts the coordinated control of leg muscles during locomotion inducing asymmetric gait patterns. However, the ability of muscle synergies and spinal maps to reflect the redistribution of the workload between legs after the trauma has not been investigated so far. Methods: To investigate this issue, twelve post-stroke and ten healthy participants were asked to walk on a treadmill at controlled speeds (0.5, 0.7, 0.9, 1.1 km/h), while the EMG activity of twelve leg muscles was recorded on both legs. The synergies underlying muscle activation and the estimated motoneuronal activity in the lumbosacral enlargement (L2-S2) were computed and compared between groups. Results: Results showed that muscle synergies in the unaffected limb were significantly more comparable to those of the healthy control group than the ones in the affected side. Spinal maps were dissimilar between the affected and unaffected sides highlighting a significant shift of the foci of the activity toward the upper levels of the spinal cord in the unaffected leg. Conclusions: Muscle synergies and spinal maps reflect the asymmetry as a motor deficit after stroke. However, further investigations are required to support or reject the hypothesis that the altered muscular organization highlighted by muscle synergies and spinal maps may be due to the concomitant contribution of the altered information coming from the upper part of the CNS, as resulting from the stroke, and to the abnormal sensory feedback due to the neuromuscular adaptation of the patients.