<p>As a notable technological advancement, archery played diverse roles in human adaptation to environmental changes and cultural interaction from the Late Pleistocene through historical periods. The identity and behavioral patterns of archery in ancient populations were closely related to their subsistence strategies, violent conflicts, ritual practices, and the organization of social and labour divisions. The ancient Subeixi people (13th–1st centuries BCE) inhabited the desert oases of the Turpan Basin, located in the central region of the Eurasian continent, where they contended with harsh environment and intricate cultural conflicts. Abundant excavated archery-related burial goods provided evidence for their widespread archery practices. We analyzed upper limb entheseal changes, biomechanical properties, and bilateral asymmetry in two Subeixi skeletal collections from the Shengjindian and Jiayi cemeteries. Our results reveal that, in the Shengjindian population, individuals buried with archery-related goods showed significantly higher EC scores at several key muscle attachment sites and reduced upper limb asymmetry, corresponding with the biomechanical patterns of archery activity. In contrast, a similar trend was not observed in the Jiayi population from an earlier period. Skeletal shape and robusticity based on geometric attributes showed no consistent or significant trends across both groups. These findings align with the hypothesis that, in ancient Subeixi society, archery was likely practiced widely as a supplementary practice in subsistence activities and interpersonal conflict, rather than as a specialized occupation. The differing patterns between the two populations further support the view that, with social development, increased cultural interactions, and changes in subsistence strategies, the role of archery may have shifted over time, with a potential trend toward specialization and intensification. </p>