Knight anoles cannot withstand cold temperatures; in winter freezes in Florida, they have been known to fall to the ground from tree canopies. In highland parts of its native Cuba, a few individuals may rest adjacent to each other during the relatively cold winters (otherwise the species is solitary and highly territorial.
In Cuba they can be seen from near sea level to an altitude of 3,300 ft in a wide range of habitats with trees, such as forest, mangrove, savanna, cultivated areas and gardens. In its introduced Florida range it even occurs in trees along roads. Knight anoles are arboreal, mostly found high in trees on the trunk or branches in the canopy, but will descend to the ground to get from one tree to another, or for thermoregulation, when occasionally seen on warm asphalt, rocks, or sidewalks.