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    Protocol for Notch-ligand Binding Assays Using Dynabeads
    Authors:  Shogo Sawaguchi, Mitsutaka Ogawa and Tetsuya Okajima, date: 10/20/2017, view: 25, Q&A: 0
    This protocol describes how to measure interaction between Notch receptors and their ligands by cell-based assay using Dynabeads. We have used the protocol to determine binding capacity between Notch1-transfected HEK293T cells and ligand-coated ...
    In vitro Co-culture of Mesenchymal Stem Cells and Endothelial Colony Forming Cells
    Authors:  Abbas Shafiee and Kiarash Khosrotehrani, date: 10/20/2017, view: 61, Q&A: 0
    The discovery of endothelial colony forming cells (ECFCs) with robust self-renewal and de novo vessel formation potentials suggests that ECFCs can be an excellent cell source for cardiovascular diseases treatment through improving neovascularization ...
    Protocol for Notch-ligand Binding Assays Using Dynabeads
    Authors:  Shogo Sawaguchi, Mitsutaka Ogawa and Tetsuya Okajima, date: 10/20/2017, view: 25, Q&A: 0
    [Abstract] This protocol describes how to measure interaction between Notch receptors and their ligands by cell-based assay using Dynabeads. We have used the protocol to determine binding capacity between Notch1-transfected HEK293T cells and ligand-coated Dynabeads. Expression of Eogt in Notch1-expressing cells promoted binding toward DLL4-coated ...
    In vitro Co-culture of Mesenchymal Stem Cells and Endothelial Colony Forming Cells
    Authors:  Abbas Shafiee and Kiarash Khosrotehrani, date: 10/20/2017, view: 61, Q&A: 0
    [Abstract] The discovery of endothelial colony forming cells (ECFCs) with robust self-renewal and de novo vessel formation potentials suggests that ECFCs can be an excellent cell source for cardiovascular diseases treatment through improving neovascularization in the ischemic tissues. However, their engraftment after transplantation resulted to be low. ...
    Labelling HaloTag Fusion Proteins with HaloTag Ligand in Living Cells
    Authors:  Huy Nguyen Duc and Xiaojun Ren , date: 09/05/2017, view: 454, Q&A: 0
    [Abstract] HaloTag has been widely used to label proteins in vitro and in vivo (Los et al., 2008). In this protocol, we describe labelling HaloTag-Cbx fusion proteins by HaloTag ligands for live-cell single-molecule imaging (Zhen et al., 2016).
    Isolation of Mouse Cardiac Neural Crest Cells and Their Differentiation into Smooth Muscle Cells
    Authors:  Xia Wang and Sophie Astrof, date: 09/05/2017, view: 376, Q&A: 0
    [Abstract] Cardiac neural crest cells (CNCCs) originate at the dorsal edge of the neural tube between the otic pit and the caudal edge of the 3rd somite, and migrate into the pharyngeal arches and the heart. We have shown that fibronectin (Fn1) plays an important role in the development of the CNCC by regulating the differentiation of CNCCs into ...
    Ex vivo Ooplasmic Extract from Developing Drosophila Oocytes for Quantitative TIRF Microscopy Analysis
    Authors:  Imre Gáspár and Anne Ephrussi, date: 07/05/2017, view: 903, Q&A: 0
    [Abstract] Understanding the dynamic behavior and the continuously changing composition of macromolecular complexes, subcellular structures and organelles is one of areas of active research in both cell and developmental biology, as these changes directly relate to function and subsequently to the development and homeostasis of the organism. Here, we ...
    Oxidative Stress Assays (arsenite and tBHP) in Caenorhabditis elegans
    Authors:  Collin Yvès Ewald, John M. Hourihan and T. Keith Blackwell, date: 07/05/2017, view: 1031, Q&A: 0
    [Abstract] Cells and organisms face constant exposure to reactive oxygen species (ROS), either from the environment or as a by-product from internal metabolic processes. To prevent cellular damage from ROS, cells have evolved detoxification mechanisms. The activation of these detoxification mechanisms and their downstream responses represent an overlapping ...
    Validating Candidate Congenital Heart Disease Genes in Drosophila
    Authors:  Jun-yi Zhu, Yulong Fu, Adam Richman and Zhe Han, date: 06/20/2017, view: 814, Q&A: 0
    [Abstract] Genomic sequencing efforts can implicate large numbers of genes and de novo mutations as potential disease risk factors. A high throughput in vivo model system to validate candidate gene association with pathology is therefore useful. We present such a system employing Drosophila to validate candidate congenital heart ...
    Physical Removal of the Midbody Remnant from Polarised Epithelial Cells Using Take-Up by Suction Pressure (TUSP)
    Authors:  Miguel Bernabé-Rubio, David C. Gershlick and Miguel A. Alonso, date: 04/20/2017, view: 1222, Q&A: 0
    [Abstract] In polarised epithelial cells the midbody forms at the apical cell surface during cytokinesis. Once severed, the midbody is inherited by one of the daughter cells remaining tethered to the apical plasma membrane where it participates in non-cytokinetic processes, such as primary ciliogenesis. Here, we describe a novel method to physically remove ...
    In vivo Mitophagy Monitoring in Caenorhabditis elegans to Determine Mitochondrial Homeostasis
    Authors:  Konstantinos Palikaras and Nektarios Tavernarakis, date: 04/05/2017, view: 1168, Q&A: 0
    [Abstract] Perturbation of mitochondrial function is a major hallmark of several pathological conditions and ageing, underlining the essential role of fine-tuned mitochondrial activity (Lopez-Otin et al., 2013). Mitochondrial selective autophagy, known as mitophagy, mediates the removal of dysfunctional and/or superfluous organelles, preserving ...
    In vivo Live Imaging of Calcium Waves and Other Cellular Processes during Fertilization in Caenorhabditis elegans
    Authors:  Jun Takayama, Masashi Fujita and Shuichi Onami, date: 04/05/2017, view: 1474, Q&A: 0
    [Abstract] Fertilization calcium waves are a conserved trigger for animal development; however, genetic analysis of these waves has been limited due to the difficulty of imaging in vivo fertilization. Here we describe a protocol to image calcium dynamics during in vivo fertilization in the genetic animal model Caenorhabditis elegans ...