Sylvilagus floridanus (Eastern Cottontail) is a common lagomorph of the eastern US. Despite a bounty of literature on the nesting ecology of this mammal, reports of arboreal nesting are largely absent from the literature. We report the first observation of an arboreal Eastern Cottontail nest since a report in 1940. The previous observation was of a nest situated in the crotch of a Salix spp. (willow) in New York; nest success was questionable. Our observation was in a suburban backyard. The rabbit placed her nest ~2.5 m above the ground, up a vertical slope of tightly-wound twining Wisteria spp. (wisteria) stems. The nest was successful; the 4 litter mates continued to use the site for ~10 days after leaving the nest before moving elsewhere in the suburban environment.