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巨噬细胞中的染色质免疫沉淀

Chromatin Immunoprecipitation in Macrophages.

作者信息

Rousselet Germain

机构信息

CEA/DRF/Jacob/iRCM/LRTS, Fontenay aux Roses, France.

INSERM U967, Fontenay aux Roses, France.

出版信息

Methods Mol Biol. 2018;1784:177-186. doi: 10.1007/978-1-4939-7837-3_17.

Abstract

Macrophages are highly polymorphic depending upon their cellular origin and their tissue environment. The different forms that a macrophage can adopt fundamentally reflect different transcription patterns. In addition, macrophages are exquisitely sensitive to a wide variety of signals coming from either infectious agents or damaged tissues. Most of the responses to these signals involve rapid and massive modifications of transcription. The control of transcription relies on the one hand on the posttranslational modification of histones, and on the other hand on the binding on the chromatin of multiple protein complexes. Immunoprecipitation of cross-linked chromatin with specific antibodies will allow to identify the DNA regions bound by the targeted protein, or carrying the targeted histone modification. By taking a snapshot of the macrophage chromatin composition, this technique will be useful to address specific macrophage biology questions at the DNA level, but also to tackle fundamental problems in transcriptional control in a highly suited model cellular system. In this chapter we describe a protocol of chromatin immunoprecipitation in murine bone marrow-derived macrophages that can easily be adapted to other macrophage populations.

摘要

巨噬细胞因其细胞起源和组织环境的不同而具有高度多态性。巨噬细胞能够呈现的不同形式从根本上反映了不同的转录模式。此外,巨噬细胞对来自感染因子或受损组织的多种信号极为敏感。对这些信号的大多数反应都涉及转录的快速而大量的修饰。转录的控制一方面依赖于组蛋白的翻译后修饰,另一方面依赖于多种蛋白质复合物与染色质的结合。用特异性抗体对交联染色质进行免疫沉淀,将有助于鉴定与靶向蛋白结合或携带靶向组蛋白修饰的DNA区域。通过对巨噬细胞染色质组成进行快照,该技术不仅有助于在DNA水平解决特定的巨噬细胞生物学问题,还能在一个非常合适的模型细胞系统中解决转录控制的基本问题。在本章中,我们描述了一种在小鼠骨髓来源的巨噬细胞中进行染色质免疫沉淀的方案,该方案可轻松适用于其他巨噬细胞群体。

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