Methods to Study Mrp4-containing Macromolecular Complexes in the Regulation of Fibroblast Migration

The overall goal of this experimental procedure is to observe the cyclic AMP dependent regulation of fibroglast migration by MRP4.High contact microscopy can help answer key questions in the field of cell biology.For example, in addition to conferring germ resistance, MRP4 can also regulate physiological process such as cell migration by modulating intracellular cyclic nucleotide concentration.The main advantage of FRET imaging that we are going to attempt today is to allow monitoring of the intracellular dynamics of cyclic AMP and cyclic AMP dependent kinase in a real-time fashion.The implication of high contact microscopy is it can be used in drug discovery.Once the system is set up, all the instrument is set up, there is no human intervention involved in this plate-based task.Though this method can provide insight into cell migration, it can be applied to other systems, such as lipe cell nonparturbing, high contact imaging of immune cell killing, cooler dilution, angiogenesis, cell proliferation, chemotaxis, and treatese phyllite.To prepare cells for high-contact microscopy under a cell culture hood, use a one percent fibronectin solution in PBS to coat the wells of a 96 well plate.Incubate the plate in a CO2 incubator at 37