Cell Biology

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    Protocols in Current Issue
    Organelle-associated rRNA Degradation
    Authors:  Jinliang Huang and Geng Wang, date: 06/05/2019, view: 329, Q&A: 0
    [Abstract] Cytosolic rRNAs are highly dynamic and can be degraded under conditions such as apoptosis, starvation and magnesium depletion. The degradation is also related to their specific localization, as fractions of cytosolic ribosomes are localized on the surfaces of intracellular organelles, such as endoplasmic reticulum (ER) and mitochondria. Such ...
    Improved Mammalian Mitochondrial RNA Isolation
    Authors:  Jinliang Huang and Geng Wang, date: 06/05/2019, view: 424, Q&A: 0
    [Abstract] Mitochondria have two sets of RNAs. One is encoded in mitochondrial genome, and the other that consists of imported RNAs within mitochondria and cytosolic RNAs associated with mitochondrial outer membrane is encoded in the nucleus. These mitochondrial RNAs play important roles in mitochondrion biosynthesis and signaling in and out of mitochondria. ...
    Quantitative Analysis of Cargo Density in Single-extracellular Vesicles by Imaging
    Authors:  Taketoshi Kajimoto and Shun-ichi Nakamura, date: 12/20/2018, view: 1234, Q&A: 0
    [Abstract] Function of extracellular vesicles such as exosomes and microvesicles is determined by their wide ranges of cargoes inside them. Even in the pure exosomes or microvesicles the cargo contents are very heterogeneous. To understand this heterogeneous nature of extracellular vesicles, we need information of the vesicles, which will give us some ...
    Detection of Internal Matrix Targeting Signal-like Sequences (iMTS-Ls) in Mitochondrial Precursor Proteins Using the TargetP Prediction Tool
    Authors:  Felix Boos, Timo Mühlhaus and Johannes M. Herrmann, date: 09/05/2018, view: 1517, Q&A: 0
    [Abstract] Mitochondria contain hundreds of proteins which are encoded by the nuclear genome and synthesized in the cytosol from where they are imported into the organelle. Sorting signals encoded in the primary and secondary sequence of these proteins mediate the recognition of newly synthesized precursor proteins and their subsequent translocation through ...
    Isolation of Intact Vacuoles from Petunia Petals and Extraction of Sequestered Glycosylated Phenylpropanoid Compounds
    Authors:  Oded Skaliter , Jasmin Ravid, Alon Cna'ani, Gony Dvir, Rafael Knafo and Alexander Vainstein, date: 07/05/2018, view: 1818, Q&A: 0
    [Abstract] Plant vacuoles are the largest compartment in plant cells, occupying more than 80% of the cell volume. A variety of proteins, sugars, pigments and other metabolites are stored in these organelles (Paris et al., 1996; Olbrich et al., 2007). Flowers produce a variety of specialized metabolites, some of which are unique to this ...
    Investigating Localization of Chimeric Transporter Proteins within Chloroplasts of Arabidopsis thaliana
    Authors:  Susumu Uehara, Yasuko Ito-Inaba and Takehito Inaba, date: 02/05/2018, view: 3064, Q&A: 0
    [Abstract] In this protocol, we describe a method to design chimeric proteins for specific targeting to the inner envelope membrane (IEM) of Arabidopsis chloroplasts and the confirmation of their localization by biochemical analysis. Specific targeting to the chloroplast IEM can be achieved by fusing the protein of interest with a transit peptide ...
    Biochemical Isolation of Myonuclei from Mouse Skeletal Muscle Tissue
    Authors:  Alicia A Cutler, Anita H Corbett and Grace K Pavlath, date: 12/20/2017, view: 4629, Q&A: 0
    [Abstract] Skeletal muscle provides the contractile force necessary for movement, swallowing, and breathing and, consequently, is necessary for survival. Skeletal muscle cells are unique in that they are extremely large cells containing thousands of nuclei. These nuclei must all work in concert to maintain skeletal muscle function and thereby maintain life. ...
    Cell-free Fluorescent Intra-Golgi Retrograde Vesicle Trafficking Assay
    Authors:  Nathanael P. Cottam and Daniel Ungar, date: 11/20/2017, view: 3090, Q&A: 0
    [Abstract] Intra-Golgi retrograde vesicle transport is used to traffic and sort resident Golgi enzymes to their appropriate cisternal locations. An assay was established to investigate the molecular details of vesicle targeting in a cell-free system. Stable cell lines were generated in which the trans-Golgi enzyme galactosyltransferase (GalT) was ...
    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: 5677, Q&A: 3
    [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 ...
    Isolation and Quantification of Plant Extracellular Vesicles
    Authors:  Brian D. Rutter, Katie L. Rutter and Roger W. Innes, date: 09/05/2017, view: 8201, Q&A: 3
    [Abstract] Extracellular vesicles (EVs) play an important role in intercellular communication by transporting proteins and RNA. While plant cells secrete EVs, they have only recently been isolated and questions regarding their biogenesis, release, uptake and function remain unanswered. Here, we present a detailed protocol for isolating EVs from the ...



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