Cell Biology

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    Protocols in Current Issue
    Bioorthogonal Labeling and Chemoselective Functionalization of Lung Extracellular Matrix
    Authors:  Zihan Ling, Yunhui Xing, Emily Reinoso Jacome, Shierly W. Fok and Xi Ren, date: 02/20/2021, view: 607, Q&A: 0

    Decellularized extracellular matrix (ECM) biomaterials derived from native tissues and organs are widely used for tissue engineering and wound repair. To boost their regenerative potential, ECM biomaterials can be functionalized via the

    ...
    Extraction and Quantification of Sphingolipids from Hemiptera Insects by Ultra-Performance Liquid Chromatography Coupled to Tandem Mass Spectrometry
    Authors:  Ni Wang, Xiaoxiao Shi, Chao Zhang, Wenwu Zhou and Zeng-Rong Zhu, date: 02/20/2021, view: 562, Q&A: 0

    Sphingolipids are major structural components of endomembranes and have also been described as an intracellular second messenger involved in various biological functions in all eukaryotes and a few prokaryotes. Ceramides (Cer), the central molecules

    ...
    Molecular and Phenotypic Characterization Following RNAi Mediated Knockdown in Drosophila
    Authors:  Saurabh Jayesh Kumar Mehta, Pradyumna A. Joshi and Ram Kumar Mishra, date: 02/20/2021, view: 641, Q&A: 0

    Loss of function studies shed significant light on the involvement of a gene or gene product in different cellular processes. Short hairpin RNA (shRNA) mediated RNA interference (RNAi) is a classical yet straightforward technique frequently used to

    ...
    An Imaging Flow Cytometry Method to Measure Citrullination of H4 Histone as a Read-out for Neutrophil Extracellular Traps Formation
    Authors:  Emilia A. Barbu, Venina M. Dominical, Laurel Mendelsohn and Swee Lay Thein, date: 02/20/2021, view: 574, Q&A: 0

    The formation of neutrophil extracellular traps (NETs) is thought to play a critical role in infections and propagating sterile inflammation. Histone citrullination is an essential and early step in NETs formation, detectable prior to the formation

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    Live Cell FRET Analysis of the Conformational Changes of Human P-glycoprotein
    Authors:  Ryota Futamata, Noriyuki Kioka and Kazumitsu Ueda, date: 02/20/2021, view: 728, Q&A: 0

    The molecular mechanisms of P-glycoprotein (P-gp; also known as MDR1 or ABCB1) have been mainly investigated using artificial membranes such as lipid-detergent mixed micelles, artificial lipid bilayers, and membrane vesicles derived from cultured

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    Live Cell FRET Analysis of the Conformational Changes of Human P-glycoprotein
    Authors:  Ryota Futamata, Noriyuki Kioka and Kazumitsu Ueda, date: 02/20/2021, view: 728, Q&A: 0
    [Abstract]

    The molecular mechanisms of P-glycoprotein (P-gp; also known as MDR1 or ABCB1) have been mainly investigated using artificial membranes such as lipid-detergent mixed micelles, artificial lipid bilayers, and membrane vesicles derived from cultured cells. Although these in vitro experiments help illustrate details about the molecular mechanisms of

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    An Imaging Flow Cytometry Method to Measure Citrullination of H4 Histone as a Read-out for Neutrophil Extracellular Traps Formation
    Authors:  Emilia A. Barbu, Venina M. Dominical, Laurel Mendelsohn and Swee Lay Thein, date: 02/20/2021, view: 574, Q&A: 0
    [Abstract]

    The formation of neutrophil extracellular traps (NETs) is thought to play a critical role in infections and propagating sterile inflammation. Histone citrullination is an essential and early step in NETs formation, detectable prior to the formation of the hallmark extracellular DNA-scaffolded strands. In addition to the classical microscopy

    ...
    Molecular and Phenotypic Characterization Following RNAi Mediated Knockdown in Drosophila
    Authors:  Saurabh Jayesh Kumar Mehta, Pradyumna A. Joshi and Ram Kumar Mishra, date: 02/20/2021, view: 641, Q&A: 0
    [Abstract]

    Loss of function studies shed significant light on the involvement of a gene or gene product in different cellular processes. Short hairpin RNA (shRNA) mediated RNA interference (RNAi) is a classical yet straightforward technique frequently used to knock down a gene for assessing its function. Similar perturbations in gene expression can be

    ...
    Extraction and Quantification of Sphingolipids from Hemiptera Insects by Ultra-Performance Liquid Chromatography Coupled to Tandem Mass Spectrometry
    Authors:  Ni Wang, Xiaoxiao Shi, Chao Zhang, Wenwu Zhou and Zeng-Rong Zhu, date: 02/20/2021, view: 562, Q&A: 0
    [Abstract]

    Sphingolipids are major structural components of endomembranes and have also been described as an intracellular second messenger involved in various biological functions in all eukaryotes and a few prokaryotes. Ceramides (Cer), the central molecules of sphingolipids, have been depicted in cell growth arrest, cell differentiation, and apoptosis.

    ...
    Bioorthogonal Labeling and Chemoselective Functionalization of Lung Extracellular Matrix
    Authors:  Zihan Ling, Yunhui Xing, Emily Reinoso Jacome, Shierly W. Fok and Xi Ren, date: 02/20/2021, view: 607, Q&A: 0
    [Abstract]

    Decellularized extracellular matrix (ECM) biomaterials derived from native tissues and organs are widely used for tissue engineering and wound repair. To boost their regenerative potential, ECM biomaterials can be functionalized via the immobilization of bioactive molecules. To enable ECM functionalization in a chemoselective manner, we have

    ...
    Rapid and Simplified Induction of Neural Stem/Progenitor Cells (NSCs/NPCs) and Neurons from Human Induced Pluripotent Stem Cells (hiPSCs)
    Authors:  Ryutaro Kajihara, Tadahiro Numakawa and Takumi Era, date: 02/05/2021, view: 266, Q&A: 0
    [Abstract]

    Human induced pluripotent stem cells (iPSCs) and their progeny displaying tissue-specific characteristics have paved the way for regenerative medicine and research in various fields such as the elucidation of the pathological mechanism of diseases and the discovery of drug candidates. iPSC-derived neurons are particularly valuable as it is

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    Immuno-electrophysiology on Neuromuscular Junctions of Drosophila Third Instar Larva
    Authors:  Raffaella Klima, Giulia Romano, Monsurat Gbadamosi, Aram Megighian and Fabian Feiguin, date: 02/05/2021, view: 213, Q&A: 0
    [Abstract]

    Alterations in synaptic transmission are critical early events in neuromuscular disorders. However, reliable methodologies to analyze the functional organization of the neuromuscular synapses are still needed. This manuscript provides a detailed protocol to analyze the molecular assembly of the neuromuscular synapses through

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    Measuring Extracellular Proton and Anionic Fluxes in Arabidopsis Pollen Tubes
    Authors:  Maria Teresa Portes and José A. Feijó, date: 02/05/2021, view: 215, Q&A: 0
    [Abstract]

    The ion-selective vibrating probe has been used to detect and quantify the magnitude and direction of transmembrane fluxes of several ions in a wide range of biological systems. Inherently non-invasive, vibrating probes have been essential to access relevant electrophysiological parameters related to apical growth and morphogenesis in pollen

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    Isolation of Murine Primary Aortic Smooth Muscle Cells
    Authors:  Max Ole Hubert, Juan Rodriguez-Vita, Lena Wiedmann and Andreas Fischer, date: 02/05/2021, view: 242, Q&A: 0
    [Abstract]

    Vascular smooth muscle cells (VSMCs) have been cultured for decades to study the role of these cells in cardiovascular disorders. The most common source of VSMCs is the rat aorta. Here we show the adaptation of this method to isolate and culture mouse aortic VSMCs. The advantage of this method is that there are many more transgenic mouse lines

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    In vitro STING Activation with the cGAMP-STINGΔTM Signaling Complex
    Authors:  Yanpu He, Celestine Hong, Darrell J. Irvine, Jiahe Li and Paula T. Hammond, date: 02/05/2021, view: 292, Q&A: 0
    [Abstract] Activating the STING (stimulator of interferon genes) signaling pathway via administration of STING agonist cyclic GMP-AMP (cGAMP) has shown great promise in cancer immunotherapy. While state-of-the-art approaches have predominantly focused on the encapsulation of cGAMP into liposomes or polymersomes for cellular delivery, we discovered that the ...



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