Jesse Brown Veterans Affairs Medical Center

3 ARTICLES PUBLISHED IN JoVE

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 Borowicz^1,2, Robert A. Winn^1,3

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.

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. Winn^2,3

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.

JoVE Journal

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. Levine^1,2,5

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.