Animals alter their behavior to cope with urbanization. Generally, urban animals show a lower antipredator response, such as short flight initiation distance (FID; distance at which animals start to flee from threat), due to fewer natural predators and habituation to human activities. Although past studies have tested the FID towards human approach in urban animals, it is not known how animals alter their FID towards natural predator with urbanization. Using the Eurasian red squirrel in Hokkaido, we investigated how they change their FID towards humans, predators (stuffed fox) and novel object (non-animal object as control), and made comparisons between urban (n=6) and rural (n=4) habitats. We found FID towards humans was shorter than other two objects in urban squirrels, while rural squirrels showed no significant difference among each object. Moreover, urban squirrels have a shorter FID in response to each object than rural squirrels. Our results suggest that urban squirrels are able to tell the risk level of human and other potential threats and change their FID accordingly. Shorter FID in urban squirrels could indicate that they become bolder towards risky situations, which may be related to a high mortality due to road-kill.