Although its importance is recently widely accepted, force measurement has been difficult in living biological systems, mainly due to the lack of the versatile noninvasive force measurement methods. The fluctuation theorem, which represents the thermodynamic properties of small fluctuating nonequilibrium systems, has been applied to the analysis of the thermodynamic properties of motor proteins in vitro. Here we extend it to the axonal transport (displacement) of endosomes. The distribution of the displacement fluctuation had three or four distinct peaks around multiples of a unit value, which the fluctuation theorem can convert into the drag force exerted on the endosomes. The results demonstrated that a single cargo vesicle is conveyed by one to three or four units of force production.