کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
5513370 1541201 2017 7 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
Live cell imaging and analysis of lipid droplets biogenesis in hepatatis C virus infected cells
موضوعات مرتبط
علوم زیستی و بیوفناوری بیوشیمی، ژنتیک و زیست شناسی مولکولی زیست شیمی
پیش نمایش صفحه اول مقاله
Live cell imaging and analysis of lipid droplets biogenesis in hepatatis C virus infected cells
چکیده انگلیسی


- A method to label, visualize and quantify LDs in living cells is presented.
- Constantly circulating low level of lipid dye are incorporated to LDs upon biogenesis.
- Lipid droplet biogenesis can be visualized under diverse experimental conditions.
- Birth of LDs co-labeled with a GFP tagged viral LD binding protein is demonstrated.

Lipid droplets (LDs) are regulated neutral lipid storage organelles having a central role in numerous cellular processes as well as in various pathologies such as metabolic disorders, immune responses and during pathogen infection. Due to the growing significance of LDs, extensive efforts are made to study the mechanism and the dynamics of their formation and life history and how are these diverted or modified by pathogens. Real-time visualization of lipid droplet biogenesis can assist in clarifying these and other important issues and may have implications towards understanding the pathogenesis of the associated diseases. Typically, LDs are post-experimentally stained using lipophilic dyes and are visualized under a microscope. Alternatively, overexpression of LD-associated proteins or immunofluorescence analyses are used to identify and follow LDs. These experimental approaches only examine a single end point of the experiment and cannot answer questions regarding LD dynamics. Here, we describe a simple and novel experimental setting that allows real-time fluorescence staining and detection of LDs in cultured living as well as infected cells. This method is quick and simple and is not restricted to a specific dye or cell line. Using this system, the biogenesis of LDs and their growth is demonstrated in cells infected with hepatitis C virus (HCV), confirming the strength of this method and the wide range of its applications.

ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Methods - Volume 127, 15 August 2017, Pages 30-36
نویسندگان
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