Cell Biology

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    Protocols in Current Issue
    Quantitative Analysis of Actin Cable Length in Yeast
    Authors:  Shane G. McInally, Jane Kondev and Bruce L. Goode, date: 05/05/2022, view: 740, Q&A: 1
    [Abstract]

    Polarized actin cables in S. cerevisiae are linear bundles of crosslinked actin filaments that are assembled by two formins, Bnr1 (localized to the bud neck), and Bni1 (localized to the bud tip). Actin is polymerized at these two sites, which results in cables extending along the cell cortex toward the back of the mother cell. These cables serve

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    Visualization and Purification of Caenorhabditis elegans Germ Granule Proteins Using Proximity Labeling
    Authors:  Hannah L. Hertz, Ian F. Price and Wen Tang, date: 04/20/2022, view: 1032, Q&A: 0
    [Abstract]

    Membraneless organelles, such as germ granules and stress granules, are liquid-like condensates formed by phase transition. Recently, we and others have adopted proximity-based labeling methods to determine the composition of these membraneless compartments. Here, we describe the use of TurboID—an engineered promiscuous biotin

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    High Throughput Blood-brain Barrier Organoid Generation and Assessment of Receptor-Mediated Antibody Transcytosis
    Authors:  Elena Kassianidou, Claire Simonneau, Alina Gavrilov and Roberto Villaseñor, date: 04/20/2022, view: 522, Q&A: 0
    [Abstract]

    Targeting receptor-mediated transcytosis (RMT) is a successful strategy for drug delivery of biologic agents across the blood-brain barrier (BBB). The recent development of human BBB organoid models is a major advancement to help characterize the mechanisms of RMT and thus accelerate the design of brain delivery technologies. BBB organoids exhibit

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    Single Molecule Tracking Nanoscopy Extended to Two Colors with MTT2col for the Analysis of Cell-Cell Interactions in Leukemia
    Authors:  Loriane Maillot, Magali Irla and Arnauld Sergé, date: 04/20/2022, view: 683, Q&A: 0
    [Abstract]

    Single molecule tracking (SMT) is a powerful technique to study molecular dynamics, and is particularly adapted to monitor the motion and interactions of cell membrane components. Assessing interactions among two molecular populations is classically performed by several approaches, including dual-color videomicroscopy, which allows monitoring of

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    In vivo Ca2+ Imaging in Mouse Salivary Glands
    Authors:  Takahiro Takano and David I. Yule, date: 04/05/2022, view: 660, Q&A: 0
    [Abstract]

    Changes in intracellular calcium drive exocrine cell activity. In the salivary gland, acetylcholine released from parasympathetic neurons mobilizes endoplasmic reticulum calcium stores in acinar cells, which consequently initiates saliva secretion. However, our understanding of the signaling cascade is mainly based on ex vivo studies

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    3D-Structured Illumination Microscopy of Centrosomes in Human Cell Lines
    [Abstract]

    The centrosome is the main microtubule-organizing center of animal cells, and is composed of two barrel-shaped microtubule-based centrioles embedded in protein dense pericentriolar material. Compositional and architectural re-organization of the centrosome drives its duplication, and enables its microtubule-organizing activity and capability to

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    Measurement of Contractile Ring Tension Using Two-photon Laser Ablation during Drosophila Cellularization
    Authors:  Swati Sharma and Richa Rikhy, date: 03/20/2022, view: 376, Q&A: 0
    [Abstract]

    Cytokinesis occurs at the final step of cell division and leads to the separation of daughter cells. It requires assembly and constriction of the actomyosin contractile ring. The phases of assembly and constriction of the contractile ring show an increase in tension in the actomyosin complex. The measurement of tension in the contractile ring is

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    Colocalization Analysis for Cryosectioned and Immunostained Tissue Samples with or without Label Retention Expansion Microscopy (LR-ExM) by JACoP
    Authors:  Xiang Zhao, Jason Garcia, Loic A. Royer and Su Guo, date: 03/05/2022, view: 1777, Q&A: 0
    [Abstract]

    Asymmetric cell division (ACD) is fundamental for balancing cell proliferation and differentiation in metazoans. During active neurogenesis in the developing zebrafish forebrain, radial glia progenitors (RGPs) mainly undergo ACD to produce one daughter with high activity of Delta/Notch signaling (proliferative cell fate) and another daughter with

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    Laser Microirradiation and Real-time Recruitment Assays Using an Engineered Biosensor
    Authors:  Carolina dos Santos Passos, Robert E. Cohen and Tingting Yao, date: 03/05/2022, view: 1516, Q&A: 0
    [Abstract]

    Double-strand breaks (DSBs) are lesions in DNA that, if not properly repaired, can cause genomic instability, oncogenesis, and cell death. Multiple chromatin posttranslational modifications (PTMs) play a role in the DNA damage response to DSBs. Among these, RNF168-mediated ubiquitination of lysines 13 or 15 at the N-terminal tail of histone H2A

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    Labeling Endogenous Proteins Using CRISPR-mediated Insertion of Exon (CRISPIE)
    Authors:  Evan A. Wilson, Tianyi Mao and Haining Zhong, date: 03/05/2022, view: 1948, Q&A: 0
    [Abstract]

    The CRISPR/Cas9 technology has transformed our ability to edit eukaryotic genomes. Despite this breakthrough, it remains challenging to precisely knock-in large DNA sequences, such as those encoding a fluorescent protein, for labeling or modifying a target protein in post-mitotic cells. Previous efforts focusing on sequence insertion to the

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