University of Debrecen 6 articles published in JoVE Cancer Research Quantifying Antibody-Dependent Cellular Cytotoxicity in a Tumor Spheroid Model: Application for Drug Discovery Isotta Sturniolo1,2, Csongor Váróczy1,3, Ákos Máté Bede1,3, Csaba Hegedűs1, Máté Á. Demény1,4, László Virág1,4 1Department of Medical Chemistry, Faculty of Medicine, University of Debrecen, 2Doctoral School of Molecular Medicine, University of Debrecen, 3National Academy of Scientist Education, 4HUN-REN-DE Cell Biology and Signaling Research Group Here, we present a method to identify compounds that modulate the ADCC mechanism, an important cancer cell-killing mechanism of antitumor antibodies. The cytotoxic effect of NK cells is measured in breast cancer cell spheroids in the presence of Trastuzumab. Image analysis identifies live and dead killer and target cells in spheroids. Cancer Research High-Content Screening Assay for the Identification of Antibody-Dependent Cellular Cytotoxicity Modifying Compounds Eliza Guti*1,2, Ákos Máté Bede*1,3, Csongor Váróczy*1,3, Csaba Hegedűs1, Máté Á. Demény1,4, László Virág1,4 1Department of Medical Chemistry, Faculty of Medicine, University of Debrecen, 2Doctoral School of Molecular Medicine, University of Debrecen, 3National Academy of Scientist Education, University of Debrecen, 4ELKH-DE Cell Biology and Signaling Research Group This protocol presents an automated, image-based high-throughput technique to identify compounds modulating natural killer cell-mediated breast cancer cell killing in the presence of a therapeutic anti-HER-2 antibody. Biology Assessing Protein Interactions in Live-Cells with FRET-Sensitized Emission György Vámosi1, Sarah Miller2, Molika Sinha2, Maria Kristha Fernandez2, Gabor Mocsár1, Malte Renz2 1Department of Biophysics and Cell Biology, Faculty of Medicine, University of Debrecen, 2Gynecologic Oncology Division, Stanford University School of Medicine Förster Resonance Energy Transfer (FRET) between two fluorophore molecules can be used for studying protein interactions in the living cell. Here, a protocol is provided as to how to measure FRET in live cells by detecting sensitized emission of the acceptor and quenching of the donor molecule using confocal laser scanning microscopy. Bioengineering Three-Dimensional Echocardiographic Method for the Visualization and Assessment of Specific Parameters of the Pulmonary Veins Csaba Jenei1, Laszlo Nagy1, Reka Urbancsek1, Daniel Czuriga1, Zoltan Csanadi1 1Division of Cardiology, Department of Cardiology, Faculty of Medicine, University of Debrecen The dimensions of the pulmonary veins (PV) are important parameters when planning pulmonary vein isolation. 2D transoesophageal echocardiography can only provide limited data about the PVs; however, 3D echocardiography can evaluate relevant diameters and areas of the PVs, as well as their spatial relationship to surrounding structures. Biochemistry Use of Recombinant Fusion Proteins in a Fluorescent Protease Assay Platform and Their In-gel Renaturation Beáta Bozóki*1,2, János András Mótyán*1, Márió Miczi1, Lívia Diána Gazda1, József Tőzsér1 1Department of Biochemistry and Molecular Biology, Faculty of Medicine, University of Debrecen, 2Biotechnology Research Department, Gedeon Richter Plc Here, we present the detailed procedure of a recently developed protease assay platform utilizing N-terminal hexahistidine/maltose-binding protein and fluorescent protein-fused recombinant substrates attached to the surface of nickel-nitrilotriacetic acid magnetic agarose beads. A subsequent in-gel analysis of the assay samples separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis is also presented. Developmental Biology Prediction and Validation of Gene Regulatory Elements Activated During Retinoic Acid Induced Embryonic Stem Cell Differentiation Zoltan Simandi*1, Attila Horvath*2, Peter Nagy1, Laszlo Nagy1,2,3 1Sanford-Burnham-Prebys Medical Discovery Institute at Lake Nona, 2Department of Biochemistry and Molecular Biology, Research Center for Molecular Medicine, Medical and Health Science Center, University of Debrecen, 3MTA-DE “Lendulet” Immunogenomics Research Group, University of Debrecen In this work we provide an experimental workflow of how active enhancers can be identified and experimentally validated.
