Reactive oxygen species (ROS) have long been considered as cytotoxic. However, recent evidence indicates a prominent role of ROS as signaling molecules in the response to hormones, growth and coagulation factors, cytokines and other factors as well as to changes in oxygen tension. The hypoxia-inducible transcription factors (HIFs) are key players in the cellular response to changes in oxygen tension. Recently, HIFs have also been shown to respond to the above-mentioned non-hypoxic stimuli. In this article, the role of ROS in the regulation of HIF-1 under hypoxic and non-hypoxic conditions is summarized.