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Variation in the size of home range of white-tailed deer (Odocoileus virginianus) has broad implications for managing populations, agricultural damage, and disease spread and transmission. Size of home range of deer also varies seasonally because plant phenology dictates the vegetation types that are used as foraging or resting sites. Knowledge of the landscape configuration and connectivity that contributes to variation in size of home range of deer for the region is needed to fully understand differences and similarities of deer ecology throughout the Midwest. We developed a research team from four Midwestern states to investigate how size of home range of deer in agro-forested landscapes is influenced by variations in landscape characteristics that provide essential habitat components. We found that for resident female deer, annual size of home range in Illinois (mean = 0.99 km2), Michigan (mean = 1.34 km2), Nebraska (mean = 1.20 km2), and Wisconsin (mean = 1.47 km2) did not differ across the region (F3,175 = 0.42, P = 0.737), but differences between agricultural growing and non-growing periods were apparent. Variables influencing size of home range included: distance to forests, roads, and urban development from the centroid of deer home range, and percent of crop as well as four landscape pattern indices (contrast-weighted edge density, mean nearest neighbor, area-weighted mean shape index, and patch size coefficient of variation). We also identified differences in model selection for four landscapes created hierarchically to reflect levels of landscape connectivity determined from perceived ability of deer to traverse the landscape. Connectivity of selected forested regions within agro-forested ecosystems across the Midwest plays a greater role in understanding the size of home ranges than traditional definitions of deer habitat conditions and landscape configuration.