26 Result(s)
within
Protocol
**an Qi
Cell Biology
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Protocol
Expression, Purification, and Nanodisc Reconstitution of Connexin-43 Hemichannels for Structural Characterization by Cryo-Electron Microscopy
Connexins are polytopic domain membrane proteins that form hexameric hemichannels (HCs) which can assemble into gap junction channels (GJCs) at the interface of two neighboring cells. The HCs may be involved i...
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High-Throughput γ-H2AX Assay Using Imaging Flow Cytometry
The γ-H2AX assay is a sensitive and reliable method to evaluate radiation-induced DNA double-strand breaks. The conventional γ-H2AX assay detects individual nuclear foci manually, but is labor-intensive and ti...
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A Surgical Approach to Hindlimb Suspension: A Mouse Model of Disuse-Induced Atrophy
Hindlimb suspension is a well-established rodent model of disuse-induced atrophy and is commonly used to simulate the effects of bed rest and space flight on humans. Over the decades, this method has undergone...
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Detection and Analysis of Microtubule Nucleator γ-Tubulin Ring Complex
Golgi-derived microtubules constitute an asymmetrical microtubule network that drives polarized transport of vesicles to support cell polarization and directional migration. Golgi-based microtubule nucleation ...
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A Fluorescence-Microscopic System for Monitoring the Turnover of the Autophagic Substrate p62/SQSTM1
In conditions of cellular stress and nutrient shortage, macroautophagy (hereafter referred to as autophagy) assures the degradation of dysfunctional macromolecules and organelles as it liberates energy resourc...
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Functional Analysis of Actin-Binding Proteins in the Central Nervous System of Drosophila
Using Drosophila actin-binding protein Dunc-115 as an example, this chapter describes a MARCM (mosaic analysis with a repressible cell marker)-based method for analyzing cytoskeletal components for their function...
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Tetramer-Associated T Cell Receptor Sequencing
Linking antigen specificity to T cell receptor (TCR) sequences is critical, albeit challenging, to both understanding T cell biology and develo** T cell-based therapeutics. Here, we describe in detail tetram...
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A Protocol for Isolation, Purification, Characterization, and Functional Dissection of Exosomes
Extracellular vesicles (EVs) are membrane-enclosed vesicles released by cells. They carry proteins, nucleic acids, and metabolites which can be transferred to a recipient cell, locally or at a distance, to eli...
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Protocols for Full Thickness Skin Wound Repair Using Prevascularized Human Mesenchymal Stem Cell Sheet
Split thickness skin grafts (STSGs) are one of the standard treatments available for full thickness wound repair when full thickness grafts (FTGs) are not viable, such as in the case of wounds with large surfa...
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Efficient Culture of Intestinal Organoids with Blebbistatin
The intestinal epithelium is one of the most rapidly self-renewing tissues throughout life in mammals. A small population of stem cells at the base of crypt in the epithelium can continually self-renew and giv...
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Full-Thickness Human Skin Equivalent Models of Atopic Dermatitis
Atopic dermatitis is a chronic inflammatory skin disease caused by complex multifactorial etiology. In the recent years, there have been significant advances in tissue engineering and the generation of in vitr...
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Genomic and Transcriptomic Analyses of Avian Sex Chromosomes and Sex-Linked Genes
Sex chromosomes and sex-linked genes usually show unusual features comparing to the rest of the genome and thus are of particular interests to evolutionary and developmental biologists. Here we describe recent...
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Probing the Role of Cyclic di-GMP Signaling Systems in Disease Using Chinese Radish
The determination of the genome sequences of pathogenic bacteria has facilitated functional analyses that aim to understand the molecular basis of virulence. In particular, genome sequence information of the p...
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Monitoring Wnt/β-Catenin Signaling in Skin
Wnt signaling through β-catenin plays a crucial role in skin development and homeostasis. Disruption or hyperactivation of this pathway results in skin defects and diseases (Lim and Nusse, Cold Spring Harb Per...
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Functional Analysis of Actin-Binding Proteins in the Central Nervous System of Drosophila
Using Drosophila actin-binding protein Dunc-115 as model system, this chapter describes a MARCM (mosaic analysis with a repressible cell marker)-based method for analyzing cytoskeletal components for their functi...
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Protocol
Assessing Reprogramming by Chimera Formation and Tetraploid Complementation
Pluripotent stem cells can be evaluated by pluripotent markers expression, embryoid body aggregation, teratoma formation, chimera contribution and even more, tetraploid complementation. Whether iPS cells in ge...
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A Chimeric Embryoid Body Model to Study Vascular Morphogenesis
Embryonic stem cell (ESC)-derived embryoid body (EB) is a unique model for studying vascular development, in that it provides a three-dimensional microenvironment that mimics an in vivo milieu. When using gene...
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Protocol
Membrane Nanotube Formation in Osteoclastogenesis
Membrane tunneling nanotubes (TNTs) are unique intercellular structures, which enable rapid transport of various materials and rapid communication between cells present in a long distance. During osteoclastoge...
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Inhibition of PI3K-Akt-mTOR Signaling in Glioblastoma by mTORC1/2 Inhibitors
Amplification of the gene encoding the epidermal growth factor receptor (EGFR) occurs commonly in glioblastoma (GBM), leading to activation of downstream kinases, including phosphatidylinositol 3′-kinase (PI3K...
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Protocol
Functional Analysis of Actin-Binding Proteins in the Central Nervous System of Drosophila
Using Drosophila actin-binding protein Dunc-115 as an example, this chapter describes an MARCM based method for analyzing cytoskeletal components for their functions in the nervous system. Following a concise des...
