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7 ARTICLES PUBLISHED IN JoVE

Environment

Transcript and Metabolite Profiling for the Evaluation of Tobacco Tree and Poplar as Feedstock for the Bio-based Industry

Colin Ruprecht¹, Takayuki Tohge¹, Alisdair Fernie¹, Cara L. Mortimer², Amanda Kozlo², Paul D. Fraser², Norma Funke¹, Igor Cesarino^3,4, Ruben Vanholme^3,4, Wout Boerjan^3,4, Kris Morreel^3,4, Ingo Burgert^5,6, Notburga Gierlinger^5,6, Vincent Bulone⁷, Vera Schneider⁸, Andrea Stockero⁸, Juan Navarro-Aviñó⁹, Frank Pudel¹⁰, Bart Tambuyser¹¹, James Hygate¹², Jon Bumstead¹³, Louis Notley¹³, Staffan Persson^1,14

¹Max Planck Institute for Molecular Plant Physiology, ²School of Biological Sciences, Plant Molecular Science, Centre for Systems and Synthetic Biology, Royal Holloway, University of London, ³Department of Plant Systems Biology, VIB, ⁴Department of Plant Biotechnology and Bioinformatics, UGhent, ⁵Institute for Building Materials, ETH Zurich, ⁶Applied Wood Materials, EMPA, ⁷Division of Glycoscience, School of Biotechnology, AlbaNova University Center, Royal Institute of Technology (KTH), ⁸European Research and Project Office GmbH, ⁹ABBA Gaia S.L., ¹⁰Pflanzenöltechnologie, ¹¹Capax Environmental Services, ¹²Green Fuels, ¹³Neutral Consulting Ltd, ¹⁴Plant Cell Biology Research Centre, School of Botany, University of Melbourne

Plant biomass offers a renewable resource for multiple products, including fuel, feed, food, and a variety of materials. In this paper we investigate the properties of tobacco tree (Nicotiana glauca) and poplar as suitable sources for a biorefinery pipeline.

Immunology and Infection

Co-immunoprecipitation of the Mouse Mx1 Protein with the Influenza A Virus Nucleoprotein

Judith Verhelst^1,2, Dorien De Vlieger^1,2, Xavier Saelens^1,2

¹Inflammation Research Center, VIB, ²Department of Biomedical Molecular Biology, Ghent University

This co-immunoprecipitation protocol allows to study the interaction between the influenza A virus nucleoprotein and the antiviral Mx1 protein in human cells. The protocol emphasizes the importance of N-ethylmaleimide for successful co-immunoprecipitation of Mx1 and influenza A virus nucleoprotein.

Immunology and Infection

Bronchoalveolar Lavage of Murine Lungs to Analyze Inflammatory Cell Infiltration

Lien Van Hoecke^1,2, Emma Richelle Job^1,2, Xavier Saelens^1,2, Kenny Roose^1,2

¹UGent Center for Medical Biotechnology, VIB, ²Department of Biomedical Molecular Biology, Ghent University (UGent)

The health status of the lung is reflected by the type and number of immune cells that are present in the bronchioles of the lung. We describe a bronchoalveolar lavage technique that allows the isolation and study of nonadherent cells and soluble factors from the lower respiratory tract of mice.

JoVE Journal

Structure-function Studies in Mouse Embryonic Stem Cells Using Recombinase-mediated Cassette Exchange

Tim Pieters^1,2,3,4, Lieven Haenebalcke^1,2, Kenneth Bruneel^1,2,4, Niels Vandamme^1,2,4, Tino Hochepied^1,2, Jolanda van Hengel⁵, Dagmar Wirth⁶, Geert Berx^1,2,4, Jody J. Haigh⁷, Frans van Roy^1,2,4, Steven Goossens^1,2,3,4

¹Department of Biomedical Molecular Biology, Ghent University, ²Inflammation Research Center, VIB, ³Center for Medical Genetics, Ghent University Hospital, ⁴Cancer Research Institute Ghent (CRIG), ⁵Department of Basic Medical Sciences, Faculty of Medicine and Health Sciences, Ghent University, ⁶Helmholtz Center for Infection Research, ⁷Mammalian Functional Genetics Laboratory, Division of Blood Cancers, Australian Centre for Blood Diseases, Department of Clinical Haematology, Monash University and Alfred Health Alfred Centre

Proteins often contain multiple domains that can exert different cellular functions. Gene knock-outs (KO) do not consider this functional diversity. Here, we report a recombination-mediated cassette exchange (RMCE)-based structure-function approach in KO embryonic stem cells that allows for the molecular dissection of various functional domains or variants of a protein.

Neuroscience

In Vivo Electrophysiological Measurement of Compound Muscle Action Potential from the Forelimbs in Mouse Models of Motor Neuron Degeneration

Eveliina Pollari^1,2, Robert Prior^1,2, Wim Robberecht^1,2,3, Philip Van Damme^1,2,3, Ludo Van Den Bosch^1,2

¹Department of Neurosciences, Experimental Neurology, KU Leuven - University of Leuven, ²Center for Brain & Disease Research, Laboratory of Neurobiology, VIB, ³Department of Neurology, University Hospitals Leuven

The measurement of nerve conduction is a useful tool to assess mouse models of neurodegeneration but it is frequently only applied to stimulate the sciatic nerve in hindlimbs. Here, we describe a technique to measure compound muscle action potential (CMAP) in vivo in the mouse forelimb muscles innervated by the brachial plexus.

Neuroscience

Generation of Human Motor Units with Functional Neuromuscular Junctions in Microfluidic Devices

Katarina Stoklund Dittlau^1,2, Emily N. Krasnow^1,2, Laura Fumagalli^1,2, Tijs Vandoorne^1,2, Pieter Baatsen^3,4, Axelle Kerstens^3,4, Giorgia Giacomazzi⁵, Benjamin Pavie^3,4, Elisabeth Rossaert^1,2, Jimmy Beckers^1,2, Maurilio Sampaolesi⁵, Philip Van Damme^1,2,6, Ludo Van Den Bosch^1,2

¹Department of Neurosciences, Experimental Neurology, and Leuven Brain Institute, KU Leuven - University of Leuven, ²VIB Bio Imaging Core, KU Leuven - University of Leuven, ³Department of Development and Regeneration, Stem Cell and Developmental Biology, KU Leuven - University of Leuven, ⁴Department of Neurology, University Hospitals Leuven

We describe a method to generate human motor units in commercially available microfluidic devices by co-culturing human induced pluripotent stem cell-derived motor neurons with human primary mesoangioblast-derived myotubes resulting in the formation of functionally active neuromuscular junctions.

Neuroscience

Microdissection and Whole Mount Scanning Electron Microscopy Visualization of Mouse Choroid Plexus

Elien Van Wonterghem^*1,2, Lien Van Hoecke^*1,2, Griet Van Imschoot^1,2, Daan Verhaege^1,2, Marlies Burgelman^1,2, Roosmarijn E. Vandenbroucke^1,2

¹VIB Center for Inflammation Research, VIB, ²Department of Biomedical Molecular Biology, Ghent University

The choroid plexus (CP), an understudied tissue in neuroscience, plays a key role in health and disease of the central nervous system. This protocol describes a microdissection technique for isolating the CP and the use of scanning electron microscopy to obtain an overall view of its cellular structure.