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Author:
Date:
2015-08-05
[Abstract] Neutrophil extracellular traps (NETs) are fibrous mesh-like, web-like, or string-like structures which are composed of DNA, histones, and granule proteins such as neutrophil elastase or myeloperoxidase. When activated by phorbol myristate acetate, interleukin-8, lipopolysaccharide (LPS), and various pathogens, neutrophils release NETs. We reported that NETs were classified as two distinct forms; cell-free NETs that were released away from neutrophils and anchored NETs that were anchored to neutrophils. In general, extracellular DNAs are used as a surrogate marker of NETs. Here, we describe a protocol regarding quantitative procedures of extracellular DNAs released from ex vivo neutrophils activated by LPS using fluorometric double-stranded DNA (dsDNA) quantification assay.
[摘要] 中性粒细胞外陷阱（NET）是由DNA，组蛋白和颗粒蛋白如嗜中性粒细胞弹性蛋白酶或髓过氧化物酶组成的纤维状网状，网状或绳状结构。 当被佛波醇肉豆蔻酸乙酯，白细胞介素-8，脂多糖（LPS）和各种病原体激活时，嗜中性粒细胞释放NETs。 我们报告，NETs被分为两种不同的形式; 从嗜中性粒细胞释放的无细胞NETs和锚定于嗜中性粒细胞的锚定NET。 通常，细胞外DNA用作NETs的替代标记。 在这里，我们描述一个议定书关于使用荧光双链DNA（dsDNA）定量测定由LPS激活的离体嗜中性粒细胞释放的细胞外DNA的定量程序。
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Author:
Date:
2014-06-20
[Abstract] Systems biology approaches can be used to study the regulatory interactions occurring between many components of the biological system at the whole-genome level and decipher the circuitries implicated in the regulation of cellular processes, including those imparting virulence to opportunistic fungi. Candida albicans (C. albicans) is a leading human fungal pathogen. It undergoes morphological switching between a budding yeast form and an elongated multicellular hyphal form. This transition is required for C. albicans’ ability to cause disease and is regulated through highly interconnected regulatory interactions between transcription factors (TFs) and target genes. The chromatin immunoprecipitation (ChIP)-High-throughput sequencing (Seq) technology (ChIP-Seq) is ...
[摘要] 系统生物学方法可以用于研究在全基因组水平上发生在生物系统的许多组分之间的调节相互作用，并且解释涉及细胞过程调节的电路，包括赋予机会性真菌的毒力的电路。 白色念珠菌（ C。albicans ）是一种主要的人类真菌病原体。它经历萌芽酵母形式和细长多细胞菌丝形式之间的形态学转换。此转换是 C所必需的。白色念珠菌引起疾病的能力，并且通过转录因子（TF）和靶基因之间高度相互关联的调节相互作用来调节。染色质免疫沉淀（ChIP） - 高通量测序（Seq）技术（ChIP-Seq）是解码转录调节网络的强有力的方法。该方案针对从丝状结构C制备ChIP DNA进行了优化。白色念珠菌细胞，然后进行高通量测序以鉴定调节酵母对菌丝转变的TF的靶标。
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