Robust locomotion relies on information from proprioceptors: sensory organs that communicate the position of body parts to the spinal cord and brain. Proprioceptive circuits in the spinal cord are known to regulate locomotion in challenging environments. Yet, the regulatory importance of the brain remains less clear. Here, we examine the role of the brain in integrating proprioceptive information during locomotion through mouse genetic studies and in vivo electrophysiology. The systemic removal of proprioceptors leaves the animals in a constantly perturbed state, similar to that observed during mechanically perturbed locomotion in wild type. In contrast, after surgical interruption of the ascending proprioceptive pathways, wild-type mice show normal walking behaviour, but lose the ability to cope with external perturbations. Our findings provide direct evidence of a pivotal role for ascending proprioceptive information in achieving safe locomotion. In this supplementary data set we made available: a) the metadata with animal information (mouse_data.xlsx); b) the raw EMG (RAW_EMG.RData); c) the touchdown and lift-off timings (CYCLE_TIMES.RData) and d) the kinematics (KINEMATICS.RData) of the recorded limb; e) the R code (muscle_synergies.R , continuously updated at https://github.com/alesantuz/musclesyneRgies) to extract muscle synergies. In total, 146 trials from 23 mice are included in the supplementary data set.