The Drosophila trachea is a premier genetic system to investigate the fundamental mechanisms of tubular organ formation. Development of the trachea consists of the well understood early branch specification and migration processes, and the less clear later branch maturation process including the apical membrane expansion, cytoskeleton rearrangement, luminal matrix clearance, and air-filling. We identified seven members of the Osiris (Osi) gene family with obvious tracheal expression in Drosophila. In addition, HA-tagged Osi proteins are highly concentrated in vesicle-like structures at and near the apical membrane. Osi proteins are predicted to contain endocytic signals and transmembrane domains. The localization of Osi proteins is consistent with these predictions. Interestingly, the Drosophila tracheal tube maturation process also occurs at the apical membrane. Taken together, the localization of Osi proteins suggest that these proteins are likely involved in tube maturation through vesicular trafficking or interacting with other apical membrane proteins.