Added PubMed Id Quantitative DIA proteomics in combination with the transcription inhibitor actinomycin D was performed on the tilapia OmB cell line to identify proteins that are upregulated by transcriptional regulation during hyperosmotic stress. This analysis revealed proteins that are transcriptionally up-regulated by hyperosmolality in these cells. The promoter regions of these proteins were compared and a novel hyperosmolality-induced cis-regulatory element (CRE) and corresponding transcription factor candidate were identified. The CRE was experimentlly validated by site-directed mutagenesis in combination with reporter assays.