Retinoblastoma (RB) tumor suppressor proteins are important regulators of the cell cycle and are implicated in a wide variety of human tumors. Genetic analysis of RB mutations in humans and in model systems indicates that individual RB proteins also have distinct functions in development. Specific target genes or mechanisms of action of individual RB proteins in developmental contexts are not well understood, however. To better understand the developmental activities of the two RB family members in Drosophila, we have characterized endogenous expression patterns of Rbf1 and Rbf2 proteins and transcripts in embryos and imaginal discs. These gene products are coexpressed at several stages of development, however, spatial and temporal differences are evident, including partly complementary patterns of expression in the embryonic central nervous system.