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Institut Curie

9 ARTICLES PUBLISHED IN JoVE

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Medicine

Revealing the Cytoskeletal Organization of Invasive Cancer Cells in 3D
Sara Geraldo 1, Anthony Simon 1, Danijela M. Vignjevic 1
1UMR 144 CNRS, Institut Curie

This article presents a method to fluorescently label collagen that can be further used for both fix and live imaging of 3D cell cultures. We also provide an optimized protocol to visualize endogenous cytoskeletal proteins of cells cultured in 3D environments.

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Bioengineering

Study of Cell Migration in Microfabricated Channels
Pablo Vargas 1, Emmanuel Terriac 2, Ana-Maria Lennon-Duménil 1, Matthieu Piel 2
1Immunité et Cancer, Institut Curie, 2Compartimentation et Dynamique Cellulaires, Institut Curie

A quantitative method to study spontaneous migration of cells in a one-dimensional confined microenvironment is described. This method takes advantage of microfabricated channels and can be used to study migration of large number of cells under different conditions in single experiments.

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Bioengineering

Stretching Micropatterned Cells on a PDMS Membrane
Nicolas Carpi 1, Matthieu Piel 1
1UMR 144, Institut Curie

This manuscript presents a technique to apply or release forces on adherent cells or tissues using unidirectional stretching.

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Developmental Biology

Protocols for Analyzing the Role of Paneth Cells in Regenerating the Murine Intestine using Conditional Cre-lox Mouse Models
Lee Parry 1, Madeleine Young 1, Fatima El Marjou 2, Alan Richard Clarke 1
1European Cancer Stem Cell Research Institute, Cardiff University, 2Institut Curie

Intestinal epithelial stem cells (ISCs) are intermingled with Paneth cells. These cells are differentiated progeny of the ISC, which support the ISCs and provide antibacterial protection. Here we demonstrate how we used transgenic conditional mouse models to establish that Paneth cells play a crucial role in maintaining the intestinal epithelia.

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Developmental Biology

Stem cell-like Xenopus Embryonic Explants to Study Early Neural Developmental Features In Vitro and In Vivo
Beatrice C. Durand 1,2,3,4
1Institut Curie, 2UMR 3387, CNRS, 3PSL Research University, 4Université Paris-Sud

In Xenopus embryos, cells from the roof of the blastocoel are pluripotent and can be programmed to generate various tissues. Here, we describe protocols to use amphibian blastocoel roof explants as an assay system to investigate key in vivo and in vitro features of early neural development.

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Bioengineering

Traction Force Microscopy to Study B Lymphocyte Activation
Anita Kumari 1, Judith Pineau 1, Ana-Maria Lennon-Duménil 1, Martial Balland 2, Paolo Pierobon 1
1Institut Curie, PSL Research University, INSERM U932, 2Laboratoire Interdisciplinaire de Physique, Université Joseph Fourier (Grenoble 1)

Here we present a protocol used to perform traction force microscopy experiments on B cells. We describe the preparation of soft polyacrylamide gels and their functionalization, as well as data acquisition at the microscope and a summary of data analysis.

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Biochemistry

Purification of Tubulin with Controlled Posttranslational Modifications and Isotypes from Limited Sources by Polymerization-Depolymerization Cycles
Satish Bodakuntla *1,2, A.S. Jijumon *1,2, Carsten Janke 1,2, Maria M. Magiera 1,2
1PSL Research University, CNRS UMR3348, Institut Curie, 2Université Paris-Saclay, CNRS UMR3348, Université Paris Sud

This protocol describes tubulin purification from small/medium-scale sources such as cultured cells or single mouse brains, using polymerization and depolymerization cycles. The purified tubulin is enriched in specific isotypes or has specific posttranslational modifications and can be used in in vitro reconstitution assays to study microtubule dynamics and interactions.

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Biology

Deciphering High-Resolution 3D Chromatin Organization via Capture Hi-C
Antonia Hauth 1, Rafael Galupa 1, Nicolas Servant 2, Laura Villacorta 1, Kai Hauschulz 3, Joke Gerarda van Bemmel 4, Agnese Loda 1, Edith Heard 1,5
1EMBL: European Molecular Biology Laboratory, 2Institut Curie, 3Agilent Technologies, 4Genmab BV, 5Collège de France

This protocol describes the Capture Hi-C method used to characterize the 3D organization of megabased-sized targeted genomic regions at high-resolution, including boundaries of topologically associating domains (TADs) and long-range chromatin interactions between regulatory and other DNA sequence elements.

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Immunology and Infection

Simultaneous Assessment of Kinship, Division Number, and Phenotype via Flow Cytometry for Hematopoietic Stem and Progenitor Cells
Alessandro Donada 1, Tamar Tak 1, Giulio Prevedello 1,2,3,4, Idan Milo 1, Ken R. Duffy 2, Leïla Perié 1
1Quantitative Immuno-hematology, CNRS UMR168, Institut Curie, 2Hamilton Institute, Maynooth University, 3Institut Curie, PSL Research University, CNRS UMR 3348, Orsay, 4Université Paris-Sud, Université Paris-Saclay, CNRS UMR 3348, Orsay

Presented here is a flow cytometry-based technique that allows for simultaneously measuring the number of cell divisions, surface cell phenotype, and cellular kinship. Those properties can be tested statistically using a permutation-based framework.

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