Hexavalent chromium (Cr(VI)) is a widely studied chemical with large amounts of data available for informing chemical mode of action (MOA). This robust knowledgebase uniquely positions Cr(VI) to serve as an important case study in developing methods to integrate high-throughput methodologies into chemical risk assessment. In vitro high-throughput screening (HTS) data from the Tox21 consortium have been generated for Cr(VI) in various cell lines. In vivo high-content screening (HCS) data have also been produced to analyze responses in the small intestine, the primary site of carcinogenesis in mice chronically exposed to high levels of Cr(VI) via drinking water.