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Protocol
Immunolabel-First-Expand-Later Expansion Microscopy Approach Using Stable STED Dyes
Multiple expansion microscopy approaches have been successfully used in the analysis of centrioles, centrosomes, and cilia, hel** to reveal the localization of numerous centrosomal and ciliary proteins at na...
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Protocol
Visualization of Cytoskeleton Organization and Dynamics in Elongating Cotton Fibers by Live-Cell Imaging
Cotton fibers are extremely elongated single cells and have long been regarded as an ideal model to investigate polarized plant cell elongation. Actin filaments (F-actin), as well as the cortical microtubules ...
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Protocol
Quantitative Analysis of Nucleoside Triphosphate Pools in Mouse Muscle Using Hydrophilic Interaction Liquid Chromatography Coupled with Tandem Mass Spectrometry Detection
Defects in deoxyribonucleoside triphosphate (dNTP) metabolism are associated with a number of mitochondrial DNA (mtDNA) depletion syndromes (MDS). These disorders affect the muscles, liver, and brain, and the ...
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Protocol
Isolation and Characterization of Brown Adipose Tissue T Cells
Brown adipose tissue (BAT) is a specialized fat depot that can dissipate energy through uncoupled respiration and thermogenesis. Various immune cells such as macrophages, eosinophils, type 2 innate lymphoid ce...
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Protocol
In Vitro Reconstitutive Base Excision Repair (BER) Assay
The mammalian cell genome is continuously exposed to endogenous and exogenous insults that modify its DNA. These modifications can be single-base lesions, bulky DNA adducts, base dimers, base alkylation, cytos...
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Protocol
Gastrointestinal and Microbiome Profiling in Rodent Models of Neurological and Psychiatric Disorders
Mice and other rodent models have been widely used to understand the role of the gut microbiome in various neurological and psychiatric disorders. Here we describe a protocol to characterize the structural and...
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Protocol
Diagnosis, Monitoring, and Prognosis of Liquid Biopsy in Cancer Immunotherapy
Liquid biopsy (LB), as a minimally invasive method of gleaning insight into the dynamics of diseases through a patient fluid sample, represents an interesting tool that can advise in disease monitoring, treatm...
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Protocol
Targeting Intracellular Antigens with pMHC-Binding Antibodies: A Phage Display Approach
Antibodies that bind peptide-MHC (pMHC) complex in a manner akin to T cell receptor (TCR) have not only helped in understanding the mechanism of TCR-pMHC interactions in the context of T cell biology but also ...
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Protocol
Immune Cell Lineage-Specific Chimerism Testing by Next-Generation Sequencing for Engraftment Monitoring After Allogeneic Hematopoietic Stem Cell Transplantation
Chimerism is an unusual state in which a person’s body comprises cells from genetically different people. Chimerism testing allows monitoring for the relative proportion of recipient and donor-derived cell sub...
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Protocol
Optochemical Control of Cell Contractility in Drosophila Embryos
Cell shape changes based on actomyosin contractility provide a driving force in tissue morphogenesis. The temporally and spatially coordinated constrictions of many cells result in changes in tissue morphology...
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Protocol
Analyzing Centrioles and Cilia by Expansion Microscopy
Expansion microscopy is an imaging method based on isotropic physical expansion of biological samples, which improves optical resolution and allows imaging of subresolutional cellular components by conventiona...
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Protocol
Development of Antiviral Vaccine Utilizing Self-Destructing Salmonella for Antigen and DNA Vaccine Delivery
Vaccines are the most effective means to prevent infectious diseases, especially for viral infection. The key to an excellent antiviral vaccine is the ability to induce long-term protective immunity against a ...
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Protocol
Method for Stimulation of Hippocampal Neurogenesis by Transient Microneedle Insertion
Brief stereotaxic insertion and removal of a microneedle into the hippocampus of mice result in stimulation of hippocampal neurogenesis. This approach has been previously applied to a mouse model of Alzheimer’...
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Protocol
Dynamic Light Scattering Analysis to Dissect Intermediates of SNARE-Mediated Membrane Fusion
Dynamic light scattering (DLS) spectroscopy provides rapid information on the size distribution of a large number of particles in a mixture. Vesicle sizes change during the merger of lipid bilayers, and DLS an...
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Protocol
iSeq: Web-Based RNA-seq Data Analysis and Visualization
Transcriptome sequencing (RNA-seq) is becoming a standard experimental methodology for genome-wide characterization and quantification of transcripts at single base-pair resolution. However, downstream analysi...
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Protocol
Targeting Intracellular Antigens with pMHC-Binding Antibodies: A Phage Display Approach
Antibodies that bind peptide-MHC (pMHC) complex in a manner akin to T-cell receptor (TCR) have not only helped in understanding the mechanism of TCR-pMHC interactions in the context of T-cell biology, but also...
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Protocol
Genetic Mouse Models: The Powerful Tools to Study Fat Tissues
Obesity and Type 2 diabetes (T2D) are associated with a variety of comorbidities that contribute to mortality around the world. Although significant effort has been expended in understanding mechanisms that mi...
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Protocol
Preparation of a Two-Photon Fluorescent Probe for Imaging H2O2 in Lysosomes in Living Cells and Tissues
Hydrogen peroxide (H2O2) plays important roles in many physiological and pathological processes. At the cellular organelle level, the abnormal concentrations of H2O2 in the lysosomes may cause redox imbalance and...
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Protocol
Generation of TCR-Like Antibodies Using Phage Display
The adaptive immune response against cancer consists of two arms: the humoral response from B cells, and the cell-mediated response from T cells. The humoral response has the advantage of diversity, theoretica...
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Protocol
Nrf2-Target Approaches in Cancer Chemoprevention Mediated by Dietary Phytochemicals
Cancer chemoprevention with natural phytochemical compounds is an emerging strategy to prevent, impede, delay, or cure cancer. This chapter reviews the basic methods used to study the cancer chemopreventive po...