Developmental Biology

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    Protocols in Current Issue
    Generation of iMyoblasts from Human Induced Pluripotent Stem Cells
    [Abstract]

    Skeletal muscle stem cells differentiated from human-induced pluripotent stem cells (hiPSCs) serve as a uniquely promising model system for investigating human myogenesis and disease pathogenesis, and for the development of gene editing and regenerative stem cell therapies. Here, we present an effective and reproducible transgene-free protocol for

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    Fluorescence Imaging of 3D Cell Models with Subcellular Resolution
    [Abstract]

    Over the past years, research has made impressive breakthroughs towards the development and implementation of 3D cell models for a wide range of applications, such as drug development and testing, organogenesis, cancer biology, and personalized medicine. Opposed to 2D cell monolayer culture systems, advanced 3D cell models better represent the in

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    ATAC-Seq of a Single Myofiber from Mus musculus
    Authors:  Korin Sahinyan, Darren M. Blackburn and Vahab D. Soleimani, date: 06/20/2022, view: 2028, Q&A: 0
    [Abstract]

    Chromatin accessibility is a key determinant of gene expression that can be altered under different physiological and disease conditions. Skeletal muscle is made up of myofibers that are highly plastic and adaptive. Therefore, assessing the genome-wide chromatin state of myofibers under various conditions is very important to gain insight into the

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    Labelling of Active Transcription Sites with Argonaute NRDE-3—Image Active Transcription Sites in vivo in Caenorhabditis elegans
    Authors:  Antoine Barrière and Vincent Bertrand, date: 06/05/2022, view: 1032, Q&A: 0
    [Abstract]

    Live labelling of active transcription sites is critical to our understanding of transcriptional dynamics. In the most widely used method, RNA sequence MS2 repeats are added to the transcript of interest, on which fluorescently tagged Major Coat Protein binds, and labels transcription sites and transcripts. Here we describe another strategy, using

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    Simple Methods for Permanent or Transient Denervation in Mouse Sciatic Nerve Injury Models
    Authors:  Alexis Osseni, Jean-Luc Thomas, Alireza Ghasemizadeh, Laurent Schaeffer and Vincent Gache, date: 06/05/2022, view: 1225, Q&A: 0
    [Abstract]

    Our ability to move and breathe requires an efficient communication between nerve and muscle that mainly takes place at the neuromuscular junctions (NMJs), a highly specialized synapse that links the axon of a motor neuron to a muscle fiber. When NMJs or axons are disrupted, the control of muscle fiber contraction is lost and muscle are paralyzed.

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    Isolation of tdTomato Expressing Inter-follicular Epidermal Melanocytes or Keratinocytes from Mouse Tail Skin
    Authors:  Samuel Pop, Oscar Urtatiz and Catherine D. Van Raamsdonk, date: 04/20/2022, view: 1755, Q&A: 0
    [Abstract]

    The epidermis is the outermost layer of the skin. It is made up of mostly keratinocytes along with a small number of melanocytes and Langerhans cells. Melanocytes produce a pigment called melanin, which is transferred to the keratinocytes, and protects these cells from damage from UV radiation, as well as generating hair and skin colours. In this

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    Isolation and Culture of Cranial Neural Crest Cells from the First Branchial Arch of Mice
    Authors:  Hiroki Ueharu, Jingwen Yang, Yoshihiro Komatsu and Yuji Mishina, date: 04/05/2022, view: 1241, Q&A: 1
    [Abstract]

    Craniofacial anomalies (CFA) are a diverse group of deformities, which affect the growth of the head and face. Dysregulation of cranial neural crest cell (NCC) migration, proliferation, differentiation, and/or cell fate specification have been reported to contribute to CFA. Understanding of the mechanisms through which cranial NCCs contribute

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    Generation, Maintenance and HBV Infection of Human Liver Organoids
    Authors:  Shahla Romal, Tanvir Hossain and Tokameh Mahmoudi, date: 03/20/2022, view: 1730, Q&A: 0
    [Abstract]

    Hepatitis B virus (HBV) infection represents a major public health problem infecting approximately 400 million people worldwide. Despite the availability of a preventive vaccine and anti-viral therapies, chronic HBV infection remains a major health issue because it increases the risk of developing liver cirrhosis and hepatocellular carcinoma

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    3D-Structured Illumination Microscopy of Centrosomes in Human Cell Lines
    [Abstract]

    The centrosome is the main microtubule-organizing center of animal cells, and is composed of two barrel-shaped microtubule-based centrioles embedded in protein dense pericentriolar material. Compositional and architectural re-organization of the centrosome drives its duplication, and enables its microtubule-organizing activity and capability to

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    NBD-lipid Uptake Assay for Mammalian Cell Lines
    Authors:  Sara Abad Herrera, Marta Grifell-Junyent and Thomas Günther Pomorski, date: 02/20/2022, view: 2258, Q&A: 0
    [Abstract]

    All eukaryotic cells are equipped with transmembrane lipid transporters, which are key players in membrane lipid asymmetry, vesicular trafficking, and membrane fusion. The link between mutations in these transporters and disease in humans highlights their essential role in cell homeostasis. Yet, many key features of their activities, their

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