University Lyon 1 7 articles published in JoVE Cancer Research Semi-automatic PD-L1 Characterization and Enumeration of Circulating Tumor Cells from Non-small Cell Lung Cancer Patients by Immunofluorescence Jessica Garcia1,2,3, David Barthelemy1,2,3, Florence Geiguer1,2, Julie Ballandier1,2,3, Kathryn W. Li4, Jean-Phillippe Aurel4, Frédérique Le Breton5, Claire Rodriguez-Lafrasse1, Brigitte Manship3, Sébastien Couraud6,7, Léa Payen1,2,3 1Laboratoire de Biochimie et Biologie Moléculaire, Groupe Hospitalier Sud, Hospices Civils de Lyon, 2Circulating Cancer (CIRCAN) Program, Hospices Civils de Lyon Cancer Institute, 3University of Lyon, Claude Bernard University, Cancer Research Center of Lyon, 4Biolidics Limited, 5Institut de Pathologie Multisites des HCL-Site Sud, Hospices Civils de Lyon, 6Acute Respiratory Disease and Thoracic Oncology Department, Lyon Sud Hospital, Hospices Civils de Lyon Cancer Institute, 7EMR-3738 Therapeutic Targeting in Oncology, Lyon Sud Medical Faculty, Lyon 1 University The characterization of circulating tumor cells (CTCs) is a popular topic in translational research. This protocol describes a semi-automatic immunofluorescence (IF) assay for PD-L1 characterization and enumeration of CTCs in non-small cell lung cancer (NSCLC) patient samples. Chemistry Analysis of Minerals Produced by hFOB 1.19 and Saos-2 Cells Using Transmission Electron Microscopy with Energy Dispersive X-ray Microanalysis Lukasz Bozycki1, Magdalena Komiazyk1, Saida Mebarek2,3,4,5,6, Rene Buchet2,3,4,5,6, Slawomir Pikula1, Agnieszka Strzelecka-Kiliszek1 1Laboratory of Lipid Biochemistry, Nencki Institute of Experimental Biology, Polish Academy of Sciences, 2Université de Lyon, 3Université Lyon 1, 4 We present a protocol to compare the state of minerals in vesicles released by two human bone cell lines: hFOB 1.19 and Saos-2. Their mineralization profiles were analyzed by Alizarin Red-S (AR-S) staining, ultraviolet (UV) light visualization, transmission electron microscopy (TEM) imaging and energy dispersive X-ray microanalysis (EDX). Biology Study of Endoplasmic Reticulum and Mitochondria Interactions by In Situ Proximity Ligation Assay in Fixed Cells Emily Tubbs1, Jennifer Rieusset2 1Lund University Diabetes Centre, Lund University, 2INSERM UMR-1060, CarMeN Laboratory, Lyon 1 University, INRA U1235, INSA of Lyon, Rockefeller and Charles Merieux Lyon-Sud Medical Universities Here, we describe a procedure to visualize and quantify with high sensitivity the endogenous interactions between the endoplasmic reticulum and mitochondria in fixed cells. The protocol features an optimized in situ proximity ligation assay targeting the inositol 1,4,5-triphosphate receptor/glucose-regulated protein 75/voltage-dependent anion channel/cyclophilin D complex at the mitochondria-associated membrane interface. Medicine Confocal Time Lapse Imaging as an Efficient Method for the Cytocompatibility Evaluation of Dental Composites Ghania Nina Attik1, Kerstin Gritsch1,2, Pierre Colon1,3, Brigitte Grosgogeat1,2 1Laboratoire des Multimatériaux et Interfaces, UMR CNRS 5615, Université Lyon1, 2UFR d'Odontologie, Université Lyon1; Service de Consultations et de Traitements Dentaires, Hospices Civils de Lyon, 3UFR d'Odontologie, Université Paris Diderot; Service d'Odontologie, APHP, Hôpital Rothschild The most studied aspect of the biocompatibility of dental composites is their cytotoxicity 3D CLSM time lapse imaging combined with fluorescent signal quantification allows sensitive evaluation of the cell fate over time. Utilizing this method could be an efficient tool to assess the cytocompatibility of dental composites. Neuroscience Culture of Isolated Floor Plate Tissue and Production of Conditioned Medium to Assess Functional Properties of Floor Plate-released Signals Camille Charoy1, Elise Arbeille1, Karine Thoinet1, Valérie Castellani1 1CGphiMC, UMR CNRS 5534, University of Lyon 1 The floor plate is a crucial structure of the developing spinal cord, providing diffusible signals to progenitors and differentiating neurons. We describe a method to produce floor-plate conditioned medium, to apply it to fresh spinal cord tissue, and to assess the consequences on proteins of interest by biochemistry. Neuroscience Detection of the Genome and Transcripts of a Persistent DNA Virus in Neuronal Tissues by Fluorescent In situ Hybridization Combined with Immunostaining Frédéric Catez1,2,5, Antoine Rousseau4, Marc Labetoulle4, Patrick Lomonte1,2,3 1Virus and Centromere Team, Centre de Génétique et Physiologie Moléculaire et Cellulaire, CNRS UMR 5534, 2Université de Lyon 1, 3Laboratoire d'excellence, LabEX DEVweCAN, 4Institut de Virologie Moléculaire et Structurale, CNRS UPR 3296, 5Centre de Recherche en Cancérologie de Lyon, INSERM U1052, CNRS UMR 5286 We established a fluorescent in situ hybridization protocol for the detection of a persistent DNA virus genome within tissue sections of animal models. This protocol enables studying infection process by codetection of the viral genome, its RNA products, and viral or cellular proteins within single cells. Medicine Microarray-based Identification of Individual HERV Loci Expression: Application to Biomarker Discovery in Prostate Cancer Philippe Pérot1,2, Valérie Cheynet1,2, Myriam Decaussin-Petrucci1,3,4, Guy Oriol1,2, Nathalie Mugnier5, Claire Rodriguez-Lafrasse1,4,6, Alain Ruffion1,4,7, François Mallet1,2 1Joint Unit Hospices de Lyon-bioMérieux, 2Medical Diagnostic Discovery Department, BioMérieux, 3Department of Pathology and Cytology, Centre Hospitalier Lyon Sud, Hospices Civils de Lyon, 4Medical Faculty, Lyon 1 University, 5Data and Knowledge Laboratory, BioMérieux, 6Department of Biochemistry and Molecular Biology, Centre Hospitalier Lyon Sud, Hospices Civils de Lyon, 7Department of Urology, Centre Hospitalier Lyon Sud, Hospices Civils de Lyon Human endogenous retroviruses (HERV), which occupy 8% of the human genome, retain scarce coding capacities but a hundred thousand long terminal repeats (LTRs). A custom Affymetrix microarray was designed to identify individual HERV locus expression and was used on prostate cancer tissues as a proof of concept for future clinical studies.