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This study's goal was to evaluate the pilot exclusive lane for motorcycles (Motovía) in Cali, Colombia, from the perception of its users in 2012–2013. A cross-sectional study using roadside surveys plus a qualitative component of individual semi-structured interviews was carried out. Road users' perceptions were investigated before and after implementation of lane dividers (LD) and were compared with other roads in the city. Perceptions were compared using chi-square tests. In the study, 293 motorcyclists, 111 cyclists, and 115 automobile drivers were interviewed. Following the installation of the LD, the majority of motorcyclists and cyclists reported that LD made easier the driving maneuvers and decreased travel time, in contrast to perceptions of automobile drivers (p < 0.001). For most motorcyclists, the Motovia was considered as safe and effective, approving its continuity and replication. Half of automobile drivers …