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Persistence of animals in urban habitats, a stark environmental contrast to natural habitats, can be explained through evaluating the mechanisms behind organism-habitat interactions. One of the most notable effects of urbanization is the change in structural habitat; vegetation is removed and modified, favoring large trees and adding artificial structures in cities, which may alter how organismal preferences for aspects of the habitat are realized. We evaluated the mechanisms by which structural habitat changes associated with urbanization alter the available vegetation and substrates on which two species of Anolis lizards perch in urban and natural forest sites in Miami, FL. We also experimentally assessed habitat preference in the lab to establish the mechanism behind habitat selection. We found that vegetation was broader in urban areas compared to natural habitats, and artificial structures in urban areas were more than twice the diameter of available natural perches. Lizards expressed their preference for broad perches by selecting broader vegetation and artificial structures compared to their availability in both habitats. With the increased availability of broad substrates in urban areas, perch diameters selected by lizards resulted in an expansion of this aspect of the structural habitat niche for both species. The two species differed, however, in other responses to altered urban habitats. Anolis cristatellus tended to avoid artificial substrates, whereas A. sagrei used both natural and artificial structures in proportion to their availabilities. This study provides a mechanistic explanation for how urbanization alters structural habitats, leading to niche expansion for organisms living in cities.