Aix Marseille Université

4 ARTICLES PUBLISHED IN JoVE

Immunology and Infection

Whole-cell MALDI-TOF Mass Spectrometry is an Accurate and Rapid Method to Analyze Different Modes of Macrophage Activation

Richard Ouedraogo¹, Aurélie Daumas^1,2, Christian Capo¹, Jean-Louis Mege¹, Julien Textoris¹

¹Unité de Recherche sur les Maladies Infectieuses Tropicales et Emergentes (URMITE), CNRS UMR 7278, INSERM U1095, Aix Marseille Université, ²Service de médecine interne, post-urgence et thérapeutique, Hôpital de la Timone

This protocol describes the use of whole-cell MALDI-TOF mass spectrometry on eukaryotic cells. Here, we illustrate the accuracy of this technique by analyzing the multiple activation states of macrophages in response to their microenvironment.

Biology

An Easy Method for Plant Polysome Profiling

Cécile Lecampion^1,2,3, Maïna Floris^1,2,3,4, Jean Raphaël Fantino^5,6, Christophe Robaglia^1,2,3, Christophe Laloi^1,2,3

¹Laboratoire de Génétique et Biophysique des Plantes, Aix-Marseille Université, ²UMR 7265 Biologie Végétale & Microbiologie Environnementales, CNRS, ³BIAM, CEA, ⁴Department of Biology, Biocenter, University of Copenhagen, ⁵Laboratoire de Chimie Bactérienne, ⁶CNRS, LCB UMR 7283, Aix Marseille Université

This protocol describes an easy method to extract and fractionate transcripts from plant tissues on the basis of the number of bound ribosomes. It allows a global estimate of translation activity and the determination of the translational status of specific mRNAs.

Neuroscience

A Rat Model of Mild Intrauterine Hypoperfusion with Microcoil Stenosis

Masahiro Tsuji¹, Jacques-Olivier Coq², Yuko Ogawa¹, Yumi Yamamoto¹, Makiko Ohshima¹

¹Department of Regenerative Medicine and Tissue Engineering, National Cerebral and Cardiovascular Center, ²Institut de Neurosciences de la Timone, CNRS, Aix Marseille Université

Mild intrauterine hypoperfusion was produced by artery stenosis with metal microcoils wrapped around the uterine and ovarian arteries in rats at embryonic day 17. This procedure produced prenatal hypoperfusion and intrauterine growth restriction.

Immunology and Infection

Gene Knock-in by CRISPR/Cas9 and Cell Sorting in Macrophage and T Cell Lines

Lichen Zhang¹, Rong Huang¹, Liaoxun Lu¹, Rui Fu¹, Guo Guo¹, Yanrong Gu¹, Zhuangzhuang Liu¹, Le He¹, Marie Malissen², Yinming Liang¹

¹The Laboratory of Genetic Regulators in the Immune System, School of Laboratory Medicine, Xinxiang Medical University, ²Centre d’Immunologie de Marseille-Luminy, Aix Marseille Université

This protocol uses fluorescent reporters and cell sorting to simplify knock-in experiments in macrophage and T cell lines. Two plasmids are used for these simplified knock-in experiments, namely a CRISPR/Cas9- and DsRed2-expressing plasmid and a homologous recombination donor plasmid expressing EBFP2, which is permanently integrated at the Rosa26 locus in immune cells.