Phagocytosis Assay of Necroptotic Cells by Cardiac Myofibroblasts
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Author:
Date:
2017-09-20
[Abstract] In myocardial infarction (MI), a plenty of cardiomyocytes undergo necrosis and necroptosis due to the lack of oxygen and nutrients. The dead cardiomyocytes are promptly engulfed by phagocytes. When the dead cells are not engulfed, the noxious contents of the cells are released outside, and thus, induce inflammation, and obstruct the function of organs. Therefore, phagocytosis is crucial for maintaining homeostasis of organs. Herein, we describe a protocol of an in vitro phagocytosis assay of necroptotic cells.
[摘要] 在心肌梗死(MI)中,由于缺氧和营养,大量心肌细胞发生坏死和坏死。 死亡心肌细胞迅速吞噬吞噬细胞。 当死细胞不被吞没时,细胞的有害物质被释放到外部,从而引起炎症,并阻碍器官的功能。 因此,吞噬对维持器官的体内平衡至关重要。 在这里,我们描述了一种神经细胞的体外吞噬作用测定方案。 【背景】以前,认为坏死性坏死细胞和坏死核细胞只能通过心脏巨噬细胞在失败的心脏中消除。 然而,我们发现负责组织纤维化的心脏肌成纤维细胞在MI后吞噬死细胞(Nakaya等,2017)。 本文中,我们提供了一种细胞凋亡细胞的体外吞噬作用的详细方案,采用在caspase-3抑制剂Z-VAD-FMK中通过TNF-α刺激进行人脑坏死的L929细胞。
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An Assay to Determine Phagocytosis of Apoptotic Cells by Cardiac Macrophages and Cardiac Myofibroblasts
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Author:
Date:
2017-09-20
[Abstract] In myocardial infarction (MI), a number of cardiomyocytes undergo apoptosis. These apoptotic cardiomyocytes are promptly engulfed by phagocytes. If the dead cells are not engulfed, their noxious contents are released outside, resulting in induction of inflammation. Therefore, the removal of these dead cells is necessary. However, the contribution of each phagocyte type to the removal of apoptotic cells in infarcted hearts remains unresolved. Here, we describe an in vitro protocol for a phagocytosis assay to compare the engulfment ability of cardiac macrophages and cardiac myofibroblasts.
[摘要] 在心肌梗死(MI)中,许多心肌细胞凋亡。 这些凋亡心肌细胞被吞噬细胞迅速吞噬。 如果死细胞没有被吞没,其有害物质被释放到外面,导致炎症的诱导。 因此,需要去除这些死细胞。 然而,每个吞噬细胞类型对梗塞心脏中凋亡细胞的去除的贡献仍未解决。 在这里,我们描述了吞噬作用测定的体外方案来比较心脏巨噬细胞和心脏肌成纤维细胞的吞噬能力。 【背景】长期以来一直认为,心脏巨噬细胞消除了失败的心脏中产生的凋亡细胞。 然而,我们发现负责组织纤维化的心脏肌成纤维细胞也具有在MI后吞噬凋亡细胞的能力(Nakaya等,2017)。 这一发现促使我们比较心脏巨噬细胞和心脏肌成纤维细胞的吞噬能力。 在这里,我们提供了一个详细的体外吞噬试验方案来评估吞噬吞噬的程度。
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Identification of RNA-binding Proteins
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Author:
Date:
2016-09-05
[Abstract] This protocol describes the extraction of RNA-binding proteins (RBPs) from cell lysates. In order to pull down target RBPs, 5-bromo-UTP (BrUTP)-incorporated RNA probes are used, which are generated by in vitro transcription. The schematic diagram (Flowchart) with procedure is indicated (Figure1 and Figure 2).
Figure 1. Schematic diagram of procedure (A-H). Flow chart of experimental procedure is indicated at A-H.
Figure 2. Linearization of plasmids by restricted enzyme. The plasmid is cut at restriction sites adjacent to its ...
[摘要] 该协议描述了从细胞裂解物中提取RNA结合蛋白(RBP)。 为了下拉靶标RBP,使用通过体外转录产生的含有5-溴-UTP(BrUTP)的RNA探针。 示意图(流程图)与过程(图1和图2)。 图1.程序示意图(AH)。实验程序的流程图在AH指示。 图2.质粒的线性化 限制性酶。在与其克隆元件相邻的限制性位点切割质粒。
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