Differentiation inducing factor (D-factor) is a recently described protein. The gene has been cloned, but little is known concerning regulation of expression of the gene. Our study showed that fibroblasts from a variety of tissues (lung, bone marrow, gingiva, foreskin) constitutively expressed D-factor RNA. Levels of expression of this gene increased in fibroblasts of each of the tissues after exposure to several stimuli including products of activated macrophages and lymphocytes (tumor necrosis factor (TNF), interleukin 1 and lymphotoxin). Other stimuli were those capable of activating either protein kinase C (12-O-tetradecanoylphorbol-13-acetate (TPA) and teleocidin), G-binding proteins (NaF) or those inhibiting protein synthesis (cycloheximide). Accumulation of D-factor RNA by TNF may in part be explained by stabilization of D-factor transcripts; TPA and cycloheximide clearly stabilized D-factor transcripts. We and others have shown that these same signals similarly stimulated fibroblasts to express RNAs coding for a variety of cytokines including three colony-stimulating factors as well as interleukins 1 and 6. Taken together, D-factor probably is a participant in the cascade of cytokines that are produced in mesenchymal cells after various stimuli such as bacterial invasion.