Examining the Kinetics of Phagocytosis-Coupled Inflammasome Activation in Murine Bone Marrow-Derived Dendritic Cells.
Examining the Kinetics of Phagocytosis-Coupled Inflammasome Activation in Murine Bone Marrow-Derived Dendritic Cells.
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检查小鼠骨髓源性树突状细胞中吞噬作用偶联的炎症小体激活的动力学。
DOI：
10.1007/978-1-0716-3338-0_20
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发表时间：
2023
期刊：
影响因子：
--
通讯作者：
Mantegazza,AdrianaR
中科院分区：
文献类型：
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作者：
Netting,DanielJ;Mantegazza,AdrianaR
在本章中，我们描述了通过免疫荧光显微镜或活细胞成像评估NLRP3和NLRC4炎性小体组装的程序，以及在吞噬作用下通过生化和免疫技术激活炎性小体的程序。我们也包括一步一步的指南，自动计数炎性小体“斑点”成像后。虽然我们的重点是小鼠骨髓来源的树突状细胞在粒细胞-巨噬细胞集落刺激因子的存在下分化，这导致细胞群类似于炎性树突状细胞，但本文描述的策略也可能适用于其他吞噬细胞。
In the present chapter, we describe procedures to assess NLRP3 and NLRC4 inflammasome assembly by immunofluorescence microscopy or live cell imaging, together with inflammasome activation by biochemical and immunological techniques upon phagocytosis. We also include a step-by-step guide to automating the counting of inflammasome “specks” after imaging. While our focus resides on murine bone marrow-derived dendritic cells differentiated in the presence of granulocyte-macrophage colony-stimulating factor, which results in a cell population that resembles inflammatory dendritic cells, the strategies described herein may apply to other phagocytes as well.