PROJECT ABSTRACT The goals of the COBRE Center for Redox Biology and Cardiovascular Disease are to establish a nationally recognized advanced research and training program in redox biology regulation of cardiovascular disease and to facilitate the development of junior faculty research programs to achieve major independent research funding. The Redox Molecular Signaling Core (Core C) provides COBRE investigators and trainees with access to modern equipment for the detection and quantification of reactive oxygen species, reactive nitrogen species, and reactive sulfur species. Additionally, this facility enables the analysis of redox-dependent control of protein function through redox proteomics and metabolomics, provides a centralized facility for cell culture hypoxia studies, and provides expertise for establishing cell culture models of redox signaling. This range of analytical techniques to assess redox signaling will be unmatched for a single facility in Louisiana. The functions of the Redox Molecular Signaling Core are accomplished through two distinct sub-cores, the Analytical Redox Biology Sub-Core and the Molecular Signaling Sub-Core. The Analytical Redox Biology Sub-Core utilizes equipment for high performance liquid chromatography (HPLC), specialized chemiluminescent detectors, and mass spectrometry (MS) to provide high quality, quantitative redox measurements at a centralized facility with dedicated technical staff. During Phase I, the institution purchased an Orbitrap Exploris 480 mass spectrometer that significantly expanded our capability to perform proteomic analysis of cell and tissue function in models of redox signaling. During Phase II, we will expand the functionality of this core through the purchase of new equipment for reactive sulfur species quantification and additional mass spectrometry equipment to enhance sensitivity. The Molecular Signaling Sub-Core offers a centralized facility for providing assistance with molecular biology, for establishing cell culture models of redox signaling, and for affording access to vital equipment for modeling hypoxic cell injury in culture and for protein analysis using high throughput, automated capillary electrophoresis. In Phase II, we will mature and expand the Molecular Signaling Sub-Core by adding additional core services based on the needs of the current COBRE faculty, such as improved methods for transient transfection of primary cells and enhanced access to next generation sequencing technologies. The faculty and staff of the Redox Molecular Signaling Core work closely with the COBRE investigators to optimize experimental design, perform analytical measurements, provide training on the use of core equipment, and assist with data collection and interpretation. These services provide an invaluable benefit to the COBRE investigators to advance their individual projects while enhancing the infrastructure of local redox-biology research.