Cell fractionation is a method used to study different cellular events like protein translocation and sequestration by disrupting cells and fractionating their contents, thus allowing an enrichment of the protein of interest. Using different concentrations of sucrose or detergent buffer formulations in combination with centrifugations, the cell fractions are separated based on their density and size.
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Evaluation of the merit of ethanolic extract of as an anti-cancer agent in human colon cancer cell lines (HCT-116).
F1000Res
January 2025
Department of Biochemistry, Kastubra Medical College Manipal, Maniapl Academy of Higher Education, Manipal, Karnataka, India.
Background: Colon cancer is the third most common cancer type worldwide. Novel alternative therapeutic anti-cancer drugs against colon cancer with less toxicity are to be explored . This study was aimed to explore the anti-proliferative and anti-migratory activity of various fractions of ethanolic leaf extract on human colon cancer cell lines (HCT-116) and to explore the potential molecular targets from the most potent plant extract fraction.
View Article and Find Full Text PDFProtocol for extraction of gut interstitial fluid in mice with two-front nutrient supply.
STAR Protoc
January 2025
Institute of Immunology and Sir Run Run Shaw Hospital, Zhejiang University School of Medicine, Hangzhou 310058, China; Liangzhu Laboratory, Zhejiang University Medical Center, Hangzhou 311113, China. Electronic address:
The intestine features a two-front nutrient supply environment, comprising an enteral side enriched with microbial and dietary metabolites and a serosal side supplied by systemic nutrients, collectively supporting intestinal and systemic homeostasis, but there is currently no optimal approach for extracting and assessing the local intestinal microenvironment. Here, we present a protocol for constructing a nutrient supply model in mice and extracting gut interstitial fluid (GIF) via centrifugation. This model and the extracted GIF are suitable for downstream analyses.
View Article and Find Full Text PDFThe Passage of Chaperonins to Extracellular Locations in Requires a Functional Dot/Icm System.
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Department of Microbiology-Immunology, Faculty of Medicine, Dalhousie University, Halifax, NS B3H 4R2, Canada.
HtpB, the chaperonin of the bacterial pathogen , is found in extracellular locations, even the cytoplasm of host cells. Although chaperonins have an essential cytoplasmic function in protein folding, HtpB exits the cytoplasm to perform extracellular virulence-related functions that support 's lifestyle. The mechanism by which HtpB reaches extracellular locations is not currently understood.
View Article and Find Full Text PDFOptimized protocol for single-cell isolation and alkaline comet assay to detect DNA damage in cells of Drosophila wing imaginal discs.
STAR Protoc
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Department of Cellular, Computational and Integrative Biology (CIBIO), University of Trento, 38123 Trento, TN, Italy; Department of Medicine, NYU Langone Medical Center, New York, NY 10016, USA. Electronic address:
Reduced expression of nucleolar genes induces stress and DNA damage. Here, we present a protocol to analyze DNA fragmentation at the single-cell level in Drosophila imaginal discs using an optimized alkaline comet assay. We describe steps for larvae development, tissue disaggregation, and single-cell dissociation.
View Article and Find Full Text PDFVirus-Mimicking Polymer Nanocomplexes Co-Assembling HCV E1E2 and Core Proteins with TLR 7/8 Agonist-Synthesis, Characterization, and In Vivo Activity.
J Funct Biomater
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Institute for Bioscience and Biotechnology Research, University of Maryland Rockville, Rockville, MD 20850, USA.
Hepatitis C virus (HCV) is a major public health concern, and the development of an effective HCV vaccine plays an important role in the effort to prevent new infections. Supramolecular co-assembly and co-presentation of the HCV envelope E1E2 heterodimer complex and core protein presents an attractive vaccine design strategy for achieving effective humoral and cellular immunity. With this objective, the two antigens were non-covalently assembled with an immunostimulant (TLR 7/8 agonist) into virus-mimicking polymer nanocomplexes (VMPNs) using a biodegradable synthetic polyphosphazene delivery vehicle.
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