When mechanical parameters of running are measured, runners have to be accustomed to testing conditions. Nevertheless, habituated runners could still show slight evolutions of their patterns at the beginning of each new running bout. This study investigated runners' stiffness adjustments during shoe and barefoot running and stiffness evolutions of shoes. Twenty-two runners performed two 4-minute bouts at 3.61 m.s-1 shod and barefoot after a 4-min warm-up period. Vertical and leg stiffness decreased during the shoe condition but remained stable in the barefoot condition, p < 0.001. Moreover, an impactor test showed that shoe stiffness increased significantly during the first 4 minutes, p < 0.001. Beyond the 4th minute, shoe properties remained stable. Even if runners were accustomed to the testing condition, as running pattern remained stable during barefoot running, they adjusted their leg and vertical stiffness during shoe running. Moreover, as measurements were taken after a 4-min warm-up period, it could be assumed that shoe properties were stable. Then the stiffness adjustment observed during shoe running might be due to further habituations of the runners to the shod condition. To conclude, it makes sense to run at least 4 minutes before taking measurements in order to avoid runners' stiffness alteration due to shoe property modifications. However, runners could still adapt to the shoe.