PROJECT The evolutionarily conserved Wnt pathway plays a critical role during metazoan development and adult stem cell maintenance. The main focus of the Lee Lab is to understand the function of the Wnt signaling pathway by deciphering its molecular mechanisms, with the ultimate goal of understanding how deregulation of the Wnt pathway can lead to Wnt-driven diseases in humans. Using biochemical methods, my lab has made breakthroughs in our fundamental understanding of Wnt signaling, including the development of the first biochemical system to study the pathway and the development of the first mathematical Wnt model (Lee-Heinrich model). The Lee Lab utilizes a high-speed floor centrifuge for preparing plasmid DNA (for transfection and transformation) and biochemical fractionation (for protein analysis and purification) to perform experiments that further the stated tasks of the parent grant, R35 GM122516. The current centrifuge is over 15 years old, and it has been serviced successfully over its lifetime until recently. Earlier in the year, the centrifuge, which is no longer built or supported by the manufacturer, broke down and could not be fixed due to a lack of available parts. My department has two similar centrifuges, but one is broken (and likely will not be fixed due to its old age); the other centrifuge is heavily used and is in another building (requiring several flights of stairs). Thus, we are requesting funds to purchase a new Beckman Coulter Avanti-JXN30 high-speed floor centrifuge to overcome these barriers. In addition, this new machine will allow us to perform sucrose density gradient centrifugation to monitor Wnt receptor activation, a feature not available on the old centrifuge.