Cytokines are immunoregulatory proteins that are secreted by T lymphocytes and other cells upon activation. A controversy exists as to whether the induction of cytokine production is mediated at the transcriptional level by the initiation of RNA synthesis or at the post-transcriptional level by the enhancement of mRNA stability. We show that in a nontransformed T cell clone the induction of all of the cytokines that are analyzed in this report is mediated transcriptionally. We also found that a constitutive signal was seen in resting cells when the probes used in the nuclear run-on assay contained some potential cross-hybridizing sequences, for example a poly (A) tract in a cDNA probe. This observation could explain the constitutive signals seen in other studies that claim cytokine production is regulated by differential mRNA stability.