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Protocol
A Method to Quantitatively Examine Heat Stress-Induced Alternative Splicing in Plants by RNA-Seq and RT-PCR
Alternative splicing (AS) of pre-mRNAs is a type of post-transcriptional regulation in eukaryotes that expands the number of mRNA isoforms. Intron retention is the primary form of AS in plants and occurs more ...
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Protocol
Reconstitution of Vesicle Budding from the TGN and Immunoisolation of Vesicles Enriched with a Specific Cargo Client
The trans-Golgi network (TGN) is an important cargo sorting station in the secretory pathway. To reveal insights into protein sorting at the TGN, it is important to develop an assay to quantify the efficiency of ...
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Protocol
Two Complementary Strategies to Quantitate Extracellular Vesicle Uptake Using Bioluminescence and Non-Lipidic Dyes
The study of the molecular mechanisms controlling extracellular vesicle uptake by a target cell is an aspect of great interest within the EV community due to EV relevance in intercellular communication for tis...
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Protocol
Quantification of Endocannabinoids from Biological Samples Using Liquid Chromatography–Tandem Mass Spectrometry
Endocannabinoids (eCBs) are a family of lipid molecules with important regulatory function in the brain and immune system. The two well-studied eCBs are arachidonic acid derivatives, N-arachidonoylethanolamine...
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Protocol
Liquid Biopsy in Adverse Neurodevelopment of Children: Problems and Prospects
Neurodevelopmental disorders in children have an important impact on the quality of life in the whole life cycle. Severe neurodevelopmental disorders will become a serious social and family burden and an impor...
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Protocol
Mass Spectrometry Approaches for Detection and Determination of Prostaglandins from Biological Samples
Accurate determination of prostaglandins (PGs) from biological samples is critical for understanding their biological functions and interactions during physiological and pathological processes. Liquid chromato...
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Protocol
Phenotypic and Functional Studies of Human Treg Cell Subpopulations
CD4+ regulatory T (Treg) cells play an important role in maintaining immune homeostasis. Although these cells were initially studied as a homogenous cohort, we now know that they have unprecedented underlying het...
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Protocol
Statistical Methods for Integrative Clustering of Multi-omics Data
Cancers are heterogeneous diseases caused by accumulated mutations or abnormal alterations at multi-levels of biological processes including genomics, epigenomics, transcriptomics, and proteomics. There is a g...
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Protocol
Aptamers as Functional Modules for DNA Nanostructures
Watson-Crick base-pairing of DNA allows the nanoscale fabrication of biocompatible synthetic nanostructures for diagnostic and therapeutic biomedical purposes. DNA nanostructure design elicits exquisite contro...
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Protocol
NIRVANA for Simultaneous Detection and Mutation Surveillance of SARS-CoV-2 and Co-infections of Multiple Respiratory Viruses
Detection and mutation surveillance of SARS-CoV-2 are crucial for combating the COVID-19 pandemic. Here we describe a lab-based method for multiplex isothermal amplification-based sequencing and real-time anal...
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Protocol
Atrial Gene Painting in Large Animal Model of Atrial Fibrillation
Gene therapy appears promising as a targeted treatment of cardiac diseases. Atrial fibrillation (AF) is the most common sustained cardiac arrhythmia and also a major contributor to stroke, heart failure, and d...
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Protocol
In Situ Observation of Abscisic Acid Distribution in Major Crop Species by Immunofluorescence Labeling
Immunofluorescent approach uses fluorophore-conjugated antibodies to target molecules of interest for the observation of their distribution and microenvironment at cellular or tissue level. In connection with ...
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Protocol
Proteomic-, Phosphoproteomic-, and Acetylomic-Based Mass Spectrometry to Identify Tissue-Specific Protein Complexes and Phosphorylation in Plant Gametogenesis
Using proteomics to analyze phosphorylation, acetylation, and other posttranslational modifications has been a very important method in biological research. Here we take the rice meiotic anther as an example t...
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Protocol
Use of FLP/FRT System to Screen for Notch Signaling Regulators in the Drosophila Wing
Mutations of genes encoding key components of the Notch signaling pathways often result in lethality at early developmental stages, making it difficult to decipher how they regulate the formation of specific c...
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Protocol
SWATH-MS Proteomic Approach to Discover Novel Protein Targets and Pathways in Response to Abscisic Acid
SWATH-MS proteomic approaches enable the identification and quantification of thousands of proteins within a single profiling experiment, which is useful for the identification of genes regulated by abscisic a...
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Protocol
Assessing In Situ Phosphoinositide–Protein Interactions Through Fluorescence Proximity Ligation Assay in Cultured Cells
Proximity ligation assay (PLA) is a well-established method for detecting in situ interactions between two epitopes with high resolution and specificity. Notably, PLA is not only a robust method for studying p...
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Protocol
Live Imaging of Epithelial-Mesenchymal Transition in Mesoderm Cells of Gastrulating Drosophila Embryos
Epithelial-mesenchymal transitions (EMTs) drive the generation of cell diversity during both evolution and development. More and more evidence has pointed to a model where EMT is not a binary switch but a reve...
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Protocol
Neighborhood Impact Factor to Study Cell-Fate Decision-Making in Cellular Communities
Cell-fate determination is a function of cell-intrinsic and -extrinsic signaling cues. Understanding the design principles governing fate control in multicellular systems remains difficult to understand and an...
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Protocol
An Immunocapture-Based Assay for Detecting Multiple Antigens in Melanoma-Derived Extracellular Vesicles
Most human cells release extracellular vesicles (EVs) of different sizes and composition, containing biomolecules characteristic from the originating tissue. In consequence, when EVs derive from a cancer cell,...
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Protocol
Generation of Chimeric Antigen Receptor T Cells Using Gammaretroviral Vectors
Manufacturing chimeric antigen receptor (CAR)-modified T cells requires incorporation of the CAR transgene, for which viral vectors are most often used. Here, we describe the generation of CAR T cells using pr...
