Max Delbruck Center for Molecular Medicine in the Helmholtz Association 5 articles published in JoVE Cancer Research Simultaneous Imaging and Flow-Cytometry-based Detection of Multiple Fluorescent Senescence Markers in Therapy-Induced Senescent Cancer Cells Eva Dovjak1, Mario Mairhofer1, Claudia Wöß1, Jingjing Qi1, Dorothy N. Y. Fan2, Clemens A. Schmitt1,2,3, Yong Yu1 1Johannes Kepler University Linz, 2Charité - University Medicine Berlin, 3Max-Delbrück-Center for Molecular Medicine in the Helmholtz Association Here we present a flow cytometry-based method for visualization and quantification of multiple senescence-associated markers in single cells. Biology Nuclei Isolation from Adult Mouse Kidney for Single-Nucleus RNA-Sequencing Janna Leiz*1,2, Christian Hinze*1,2, Anastasiya Boltengagen3, Caroline Braeuning4, Christine Kocks3, Nikolaus Rajewsky3, Kai M. Schmidt-Ott1,2 1Department of Nephrology and Medical Intensive Care Medicine, Charité - Universitätsmedizin Berlin, corporate member of Freie Universität Berlin and Humboldt-Universität zu Berlin, 2Molecular and Translational Kidney Research, Max-Delbrück-Center for Molecular Medicine in the Helmholtz Association (MDC), 3Berlin Institute of Medical Systems Biology (BIMSB), Systems Biology of Gene Regulatory Elements, Max-Delbrück-Center for Molecular Medicine in the Helmholtz Association (MDC), 4Berlin Institute of Medical Systems Biology (BIMSB), Max Delbrück Center for Molecular Medicine in the Helmholtz Association (MDC) Here, we present a protocol to isolate high-quality nuclei from frozen mouse kidneys that improve the representation of medullary kidney cell types and avoids the gene expression artifacts from enzymatic tissue dissociation. Medicine Magnetic Resonance Imaging of Multiple Sclerosis at 7.0 Tesla Sonia Waiczies*1, Antje Els*1, Joseph Kuchling*2,3,4, Karin Markenroth Bloch5, Anna Pankowska6,7, Helmar Waiczies8, Carl Herrmann1, Claudia Chien2,3, Carsten Finke4,9, Friedemann Paul2,3,4, Thoralf Niendorf1,2,8 1Berlin Ultrahigh Field Facility (B.U.F.F.), Max Delbrück Center for Molecular Medicine in the Helmholtz Association, 2Experimental and Clinical Research Center, Max Delbrueck Center for Molecular Medicine and Charité – Universitätsmedizin Berlin, corporate member of Freie Universität Berlin, Humboldt-Universität zu Berlin, and Berlin Institute of Health, 3NeuroCure Clinical Research Center, Charité – Universitätsmedizin Berlin, corporate member of Freie Universität Berlin, Humboldt-Universität zu Berlin, and Berlin Institute of Health, 4Department of Neurology, Charité – Universitätsmedizin Berlin, corporate member of Freie Universität Berlin, Humboldt-Universität zu Berlin, and Berlin Institute of Health, 5The Swedish National 7T Facility, Lund University Bioimaging Center, Lund University, 6Department of Radiography, Medical University of Lublin, 7ECOTECH-COMPLEX, Maria Curie-Skłodowska University, 8MRI.TOOLS GmbH, 9Berlin School of Mind and Brain, Humboldt-Universität zu Berlin Here, we present a protocol to acquire magnetic resonance (MR) images of multiple sclerosis (MS) patient brains at 7.0 Tesla. The protocol includes preparation of the setup including the radio-frequency coils, standardized interview procedures with MS patients, subject positioning in the MR scanner and MR data acquisition. Medicine Electroporation-Based Genetic Modification of Primary Human Pigment Epithelial Cells Using the Sleeping Beauty Transposon System Sandra Johnen1, Nina Harmening2,3, Corinne Marie4,5, Daniel Scherman4, Zsuzsanna Izsvák6, Zoltán Ivics7, Peter Walter1, Gabriele Thumann2,3 1Department of Ophthalmology, University Hospital RWTH Aachen, 2Experimental Ophthalmology, University of Geneva, 3Department of Ophthalmology, University Hospitals of Geneva, 4Université de Paris, CNRS, INSERM, UTCBS, Unité des technologies Chimiques et Biologiques pour la Santé, 5Chimie ParisTech, PSL Research University, 6Max Delbrück Center for Molecular Medicine in the Helmholtz Association, 7Division of Medical Biotechnology, Paul-Ehrlich-Institute We have developed a protocol to transfect primary human pigment epithelial cells by electroporation with the gene encoding pigment epithelium-derived factor (PEDF) using the Sleeping Beauty (SB) transposon system. Successful transfection was demonstrated by quantitative polymerase chain reaction (qPCR), immunoblotting, and enzyme-linked immunosorbent assay (ELISA). Developmental Biology Application of RNAi and Heat-shock-induced Transcription Factor Expression to Reprogram Germ Cells to Neurons in C. elegans Ena Kolundzic1, Stefanie Seelk1, Baris Tursun1 1Berlin Institute for Medical Systems Biology, Max Delbrück Center for Molecular Medicine in the Helmholtz Association This protocol describes how to study cellular processes during cell fate conversion in Caenorhabditis elegans in vivo. Using transgenic animals, allowing heat-shock promoter-driven overexpression of the neuron fate-inducing transcription factor CHE-1 and RNAi-mediated depletion of the chromatin-regulating factor LIN-53 germ cell to neuron reprogramming can be observed in vivo.