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    Protocols in Current Issue
    Cytosolic and Nuclear Delivery of CRISPR/Cas9-ribonucleoprotein for Gene Editing Using Arginine Functionalized Gold Nanoparticles
    Authors:  Rubul Mout and Vincent M. Rotello, date: 10/20/2017, view: 34, Q&A: 0
    In this protocol, engineered Cas9-ribonucleoprotein (Cas9 protein and sgRNA, together called Cas9-RNP) and gold nanoparticles are used to make nanoassemblies that are employed to deliver Cas9-RNP into cell cytoplasm and nucleus. Cas9 protein is ...
    Labeling Aversive Memory Trace in Mouse Using a Doxycycline-inducible Expression System
    Authors:  Jianyang Du and Erin E. Koffman, date: 10/20/2017, view: 32, Q&A: 0
    A memory trace, also known as a memory engram, is theorized to be a mechanism for physical memory storage in the brain (Silva et al., 2009; Josselyn, 2010) and memory trace is associated with a specific population of neurons (Liu et al. ...
    Histochemical Preparations to Depict the Structure of Cauliflower Leaf Hydathodes
    Authors:  Aude Cerutti, Marie-Christine Auriac, Laurent D. Noël and Alain Jauneau, date: 10/20/2017, view: 19, Q&A: 0
    Hydathodes are plant organs present on leaf margins of a wide range of vascular plants and are the sites of guttation. Both anatomy and physiology of hydathodes are poorly documented. We have recently reported on the anatomy of cauliflower and ...
    Capturing Z-stacked Confocal Images of Living Bacteria Entering Hydathode Pores of Cauliflower
    Authors:  Aude Cerutti and Alain Jauneau, date: 10/20/2017, view: 31, Q&A: 0
    The present protocol to visualize living bacteria at the pore level of cauliflower hydathodes is simple and trained users in confocal microscopy can execute it successfully. It can be easily adapted to capture images with other plant-microorganism ...
    Isolation and Purification of Schwann Cells from Spinal Nerves of Neonatal Rat
    Authors:  Jinkun Wen, Dandan Tan, Lixia Li and Jiasong Guo, date: 10/20/2017, view: 38, Q&A: 0
    Primary cultured Schwann cells (SCs) are widely used in the investigation of the biology of SC and are important seed cells for neural tissue engineering. Here, we describe a novel protocol for harvesting primary cultured SCs from neonatal ...
    Establishing a Symbiotic Interface between Cultured Ectomycorrhizal Fungi and Plants to Follow Fungal Phosphate Metabolism
    In ectomycorrhizal plants, the fungal cells colonize the roots of their host plant to create new organs called ectomycorrhizae. In these new organs, the fungal cells colonize the walls of the cortical cells, bathing in the same apoplasm as the plant ...
    A Method for Radioactive Labelling of Hebeloma cylindrosporum to Study Plant-fungus Interactions
    In order to quantify P accumulation and P efflux in the ectomycorrhizal basidiomycete fungus Hebeloma cylindrosporum, we supplied 32P to mycelia previously grown in vitro in liquid medium. The culture had four main steps ...
    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 ...
    NP-40 Fractionation and Nucleic Acid Extraction in Mammalian Cells
    Authors:  Alvaro E. Galvis, Hugh E. Fisher and David Camerini, date: 10/20/2017, view: 92, Q&A: 0
    This technique allows for efficient, highly purified cytoplasmic and nuclear-associated compartment fractionation utilizing NP-40 detergent in mammalian cells. The nuclear membrane is not disturbed during the fractionation thus leaving all nuclear ...
    Cytosolic and Nuclear Delivery of CRISPR/Cas9-ribonucleoprotein for Gene Editing Using Arginine Functionalized Gold Nanoparticles
    Authors:  Rubul Mout and Vincent M. Rotello, date: 10/20/2017, view: 34, Q&A: 0
    [Abstract] In this protocol, engineered Cas9-ribonucleoprotein (Cas9 protein and sgRNA, together called Cas9-RNP) and gold nanoparticles are used to make nanoassemblies that are employed to deliver Cas9-RNP into cell cytoplasm and nucleus. Cas9 protein is engineered with an N-terminus glutamic acid tag (E-tag or En, where n = the number of glutamic acid in ...
    Labeling Aversive Memory Trace in Mouse Using a Doxycycline-inducible Expression System
    Authors:  Jianyang Du and Erin E. Koffman, date: 10/20/2017, view: 32, Q&A: 0
    [Abstract] A memory trace, also known as a memory engram, is theorized to be a mechanism for physical memory storage in the brain (Silva et al., 2009; Josselyn, 2010) and memory trace is associated with a specific population of neurons (Liu et al., 2012; Ramirez et al., 2013). Labeling and stimulating those neurons will activate ...
    Histochemical Preparations to Depict the Structure of Cauliflower Leaf Hydathodes
    Authors:  Aude Cerutti, Marie-Christine Auriac, Laurent D. Noël and Alain Jauneau, date: 10/20/2017, view: 19, Q&A: 0
    [Abstract] Hydathodes are plant organs present on leaf margins of a wide range of vascular plants and are the sites of guttation. Both anatomy and physiology of hydathodes are poorly documented. We have recently reported on the anatomy of cauliflower and Arabidopsis thaliana hydathodes and on their infection by the vascular pathogenic bacterium ...
    Capturing Z-stacked Confocal Images of Living Bacteria Entering Hydathode Pores of Cauliflower
    Authors:  Aude Cerutti and Alain Jauneau, date: 10/20/2017, view: 31, Q&A: 0
    [Abstract] The present protocol to visualize living bacteria at the pore level of cauliflower hydathodes is simple and trained users in confocal microscopy can execute it successfully. It can be easily adapted to capture images with other plant-microorganism interactions at the leaf surface and should be useful to obtain important information on pore and ...
    Isolation and Purification of Schwann Cells from Spinal Nerves of Neonatal Rat
    Authors:  Jinkun Wen, Dandan Tan, Lixia Li and Jiasong Guo, date: 10/20/2017, view: 38, Q&A: 0
    [Abstract] Primary cultured Schwann cells (SCs) are widely used in the investigation of the biology of SC and are important seed cells for neural tissue engineering. Here, we describe a novel protocol for harvesting primary cultured SCs from neonatal Sprague-Dawley (SD) rats. In the present protocol, dissociated SCs are isolated from the spinal nerves of ...
    Establishing a Symbiotic Interface between Cultured Ectomycorrhizal Fungi and Plants to Follow Fungal Phosphate Metabolism
    [Abstract] In ectomycorrhizal plants, the fungal cells colonize the roots of their host plant to create new organs called ectomycorrhizae. In these new organs, the fungal cells colonize the walls of the cortical cells, bathing in the same apoplasm as the plant cells in a space named the ‘Hartig net’, where exchanges between the two partners take place. ...
    A Method for Radioactive Labelling of Hebeloma cylindrosporum to Study Plant-fungus Interactions
    [Abstract] In order to quantify P accumulation and P efflux in the ectomycorrhizal basidiomycete fungus Hebeloma cylindrosporum, we supplied 32P to mycelia previously grown in vitro in liquid medium. The culture had four main steps that are 1) growing the mycelium on complete medium with P, 2) transfer the mycelia into new ...
    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. ...
    NP-40 Fractionation and Nucleic Acid Extraction in Mammalian Cells
    Authors:  Alvaro E. Galvis, Hugh E. Fisher and David Camerini, date: 10/20/2017, view: 92, Q&A: 0
    [Abstract] This technique allows for efficient, highly purified cytoplasmic and nuclear-associated compartment fractionation utilizing NP-40 detergent in mammalian cells. The nuclear membrane is not disturbed during the fractionation thus leaving all nuclear and perinuclear associated components in the nuclear fraction. This protocol has been modified from ...
    DQ-Red BSA Trafficking Assay in Cultured Cells to Assess Cargo Delivery to Lysosomes
    Authors:  Rituraj Marwaha and Mahak Sharma, date: 10/05/2017, view: 774, Q&A: 0
    [Abstract] Lysosomes are the terminal end of the endocytic pathway having acidic environment required for active hydrolases that degrade the cargo delivered to these compartments. This process of cargo delivery and degradation by endo-lysosomes is a tightly regulated process and important for maintaining cellular homeostasis. Cargos like EGF (Epidermal ...