Cell Biology

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    Protocols in Current Issue
    Ex vivo Assessment of Mitochondrial Function in Human Peripheral Blood Mononuclear Cells Using XF Analyzer
    Authors:  Alica Schöller-Mann, Katja Matt, Barbara Hochecker and Jörg Bergemann, date: 04/05/2021, view: 121, Q&A: 0
    [Abstract]

    Cellular health and function, as we know today, depend on a large extent on mitochondrial function. The essential function of mitochondria is the energy production, more precisely ATP production, via oxidative phosphorylation. Mitochondrial energy production parameters therefore represent important biomarkers. Studies on human cells have mainly

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    A Spectrofluorophotometrical Method Based on Fura-2-AM Probe to Determine Cytosolic Ca2+ Level in Pseudomonas syringae Complex Bacterial Cells
    [Abstract]

    Calcium signaling is an emerging mechanism by which bacteria respond to environmental cues. To measure the intracellular free-calcium concentration in bacterial cells, [Ca2+]i, a simple spectrofluorometric method based on the chemical probe Fura 2-acetoxy methyl ester (Fura 2-AM) is here presented using Pseudomonad bacterial cells. This is an

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    Liposomal Clodronate-mediated Macrophage Depletion in the Zebrafish Model
    Authors:  Linlin Yang, Alison M. Rojas and Celia E. Shiau, date: 03/20/2021, view: 579, Q&A: 0
    [Abstract]

    The ability to conduct in vivo macrophage-specific depletion remains an effective means to uncover functions of macrophages in a wide range of physiological contexts. Compared to the murine model, zebrafish offer superior imaging capabilities due to their optical transparency starting from a single-cell stage to throughout larval development.

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    Real-time Base Excision Repair Assay to Measure the Activity of the 8-oxoguanine DNA Glycosylase 1 in Isolated Mitochondria of Human Skin Fibroblasts
    Authors:  Daniel Schniertshauer, Daniel Gebhard and Jörg Bergemann, date: 03/20/2021, view: 484, Q&A: 0
    [Abstract]

    7,8-dihydro-8-oxoguanine (8-oxoG) is one of the most common and mutagenic oxidative DNA damages induced by reactive oxygen species (ROS). Since ROS is mainly produced in the inner membranes of the mitochondria, these organelles and especially the mitochondrial DNA (mtDNA) contained therein are particularly affected by this damage. Insufficient

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    Assay for Assessing Mucin Binding to Bacteria and Bacterial Proteins
    [Abstract]

    Legionella pneumophila, a Gram-negative bacterium and the causative agent of Legionnaires’ disease, exports over 300 effector proteins/virulence factors, through its type II (T2SS) and type IV secretion systems (T4SS). One such T2SS virulence factor, ChiA, not only functions as a chitinase, but also as a novel mucinase, which we believe aids

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    Generation and Implementation of Reporter BHK-21 Cells for Live Imaging of Flavivirus Infection
    Authors:  Jorge L. Arias-Arias and Rodrigo Mora-Rodríguez, date: 03/05/2021, view: 452, Q&A: 0
    [Abstract]

    The genus Flavivirus within the family Flaviviridae includes many viral species of medical importance, such as yellow fever virus (YFV), Zika virus (ZIKV), and dengue virus (DENV), among others. Presently, the identification of flavivirus-infected cells is based on either the immunolabeling of viral proteins, the application of recombinant

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    Carboxyfluorescein Dye Uptake to Measure Connexin-mediated Hemichannel Activity in Cultured Cells
    [Abstract]

    Connexins are membrane bound proteins that facilitate direct and local paracrine mediated cell-to-cell communication through their ability to oligomerise into hexameric hemichannels. When neighbouring channels align, they form gap-junctions that provide a direct route for information transfer between cells. In contrast to intact gap junctions,

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    Induction of Epithelial-mesenchymal Transition in MDCK II Cells
    Authors:  Agnieszka Pastuła and Richard Lundmark, date: 02/05/2021, view: 432, Q&A: 0
    [Abstract]

    Epithelial-mesenchymal transition (EMT) is a reversible process of epithelial cell transdifferentiation into a mesenchymal cell, that enables initiation of cell migration. EMT plays an important role in embryonic development, tissue repair and cancer metastasis. Better understanding of cellular and molecular events during EMT will not only provide

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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: 425, 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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    Multiplex T-cell Stimulation Assay Utilizing a T-cell Activation Reporter-based Detection System
    Authors:  Laurie G. Landry, Sarah E. Mann, Amanda M. Anderson and Maki Nakayama, date: 01/20/2021, view: 1118, Q&A: 0
    [Abstract]

    Immune tolerance and response are both largely driven by the interactions between the major histocompatibility complex (MHC) expressed by antigen presenting cells (APCs), T-cell receptors (TCRs) on T-cells, and their cognate antigens. Disordered interactions cause the pathogenesis of autoimmune diseases such as type 1 diabetes. Therefore, the

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