Jesse Brown VA Medical Center 5 articles published in JoVE Biology Methods to Study Epithelial Transport Protein Function and Expression in Native Intestine and Caco-2 Cells Grown in 3D Arivarasu N. Anabazhagan*1, Ishita Chatterjee*1, Shubha Priyamvada1, Anoop Kumar1, Sangeeta Tyagi1, Seema Saksena1,2, Waddah A. Alrefai1,2, Pradeep K. Dudeja1,2, Ravinder K. Gill1 1Department of Medicine, University of Illinois at Chicago, 2Department of Research, Jesse Brown VA Medical Center We describe simple methods to study the regulation of intestinal serotonin transporter (SERT) function and expression using an in vitro cell culture model of Caco-2 cells grown in 3D and an ex vivo model of mouse intestines. These methods are applicable to the study of other epithelial transporters. Biology Identification of Intracellular Signaling Events Induced in Viable Cells by Interaction with Neighboring Cells Undergoing Apoptotic Cell Death Snezana Vujicic*1,2, Lanfei Feng*1,2, Angelika Antoni3, Joyce Rauch4, Jerrold S. Levine1,2,5 1Section of Nephrology, Department of Medicine, University of Illinois at Chicago, 2Section of Nephrology, Department of Medicine, Jesse Brown Veterans Affairs Medical Center, 3Department of Biology, Kutztown University of Pennsylvania, 4Division of Rheumatology, Department of Medicine, Research Institute of the McGill University Health Centre, 5Department of Microbiology & Immunology, University of Illinois at Chicago Here, we present a protocol for the determination of intracellular signaling events induced in viable cells by physical interaction with adjacent dead or dying cells. The protocol focuses on signaling events induced by receptor-mediated recognition of the dead cells, as opposed to their phagocytic uptake or release of soluble mediators. Bioengineering Epithelial Cell Repopulation and Preparation of Rodent Extracellular Matrix Scaffolds for Renal Tissue Development Joseph S. Uzarski1,2, Jimmy Su1,2,3,4, Yan Xie1,2, Zheng J. Zhang1,2, Heather H. Ward5, Angela Wandinger-Ness6, William M. Miller7,8, Jason A. Wertheim1,2,3,4,8,9 1Comprehensive Transplant Center, Feinberg School of Medicine, Northwestern University, 2Department of Surgery, Feinberg School of Medicine, Northwestern University, 3Department of Biomedical Engineering, Northwestern University, 4Simpson Querrey Institute for BioNanotechnology in Medicine, Northwestern University, 5Department of Internal Medicine, University of New Mexico HSC, 6Department of Pathology, University of New Mexico HSC, 7Department of Chemical and Biological Engineering, Northwestern University, 8Chemistry of Life Processes Institute, Northwestern University, 9Department of Surgery, Jesse Brown VA Medical Center This protocol describes decellularization of Sprague Dawley rat kidneys by antegrade perfusion of detergents through the vasculature, producing acellular renal extracellular matrices that serve as templates for repopulation with human renal epithelial cells. Recellularization and use of the resazurin perfusion assay to monitor growth is performed within specially-designed perfusion bioreactors. Biology The Soft Agar Colony Formation Assay Stanley Borowicz1, Michelle Van Scoyk2, Sreedevi Avasarala2, Manoj Kumar Karuppusamy Rathinam2, Jordi Tauler2, Rama Kamesh Bikkavilli2, Robert A. Winn2,3 1Department of Hematology and Oncology, University of Illinois at Chicago, 2Department of Pulmonary, Critical Care, Sleep, and Allergy, University of Illinois at Chicago, 3Jesse Brown Veterans Affairs Medical Center The soft agar colony formation assay is a method used to confirm cellular anchorage-independent growth in vitro. The goal of this protocol is to illustrate a stringent method for the detection of the tumorigenic potential of transformed cells and the tumor suppressive effects of proteins on transformed cells. Biology In vitro Methylation Assay to Study Protein Arginine Methylation Rama Kamesh Bikkavilli1, Sreedevi Avasarala1, Michelle Van Scoyk1, Manoj Kumar Karuppusamy Rathinam1, Jordi Tauler1, Stanley Borowicz1,2, Robert A. Winn1,3 1Department of Pulmonary, Critical Care, Sleep, and Allergy, University of Illinois at Chicago, 2Department of Hematology and Oncology, University of Illinois at Chicago, 3Jesse Brown Veterans Affairs Medical Center Protein arginine methylation, catalyzed by a class of enzymes viz., protein arginine methyl transferases (PRMTs), is the process of enzymatic addition of methyl group(s) to arginines within proteins. The in vitro methylation assay is the most dependable tool for assessing the methylation status of known or novel PRMT substrates.