Fast detection of ambient danger is crucial for the survival of biological entities. Previous studies have shown that threatening information can bias human visual perception and enhance physiological reactions. It remains to be delineated whether the modulation of threat on human perceptual and physiological responses can take place below awareness. To probe this issue, we adopted visual looming stimuli and created two levels of threat by varying their motion trajectories to the observers, such that the stimuli could move in a path that either collided with the observers heads or just nearly missed. We found that when the observers could not explicitly discriminate any difference between the collision and the near-miss stimuli, the visual stimuli on the collision course appeared larger and evoked greater pupil constrictions than those on the near-miss course. Furthermore, the magnitude of size overestimation was comparable to when the impending collision was consciously perceived. Our findings suggest that threatening information can bias human visual perception and strengthen pupil constrictions independent of conscious representation of the threat, and imply the existence of the subcortical visual pathway dedicated to automatically processing threat-related signals in humans.