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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Preparation of Knockdown Transformants of Unicellular Charophycean Alga, Closterium peracerosum-strigosum-littorale Complex
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Author:
Date:
2016-05-20
[Abstract] To prepare the knockdown transformants of the Closterium peracerosum-strigosum-littorale (C. psl.) complex, particle bombardment was applied with a newly constructed vector (pSA0104) with an endogenous constitutive promoter fused to a DNA fragment corresponding to an antisense strand of a target gene. Using a hygromycin resistance gene (aph7”), hygromycin-resistant colonies were selected. After the second screening, integration of the vector into the genome was checked by PCR and the knockdown effect was evaluated by Western blotting using a specific antibody against the target protein.
[摘要] 为了制备鬼臼藓藻(Clonsterium peracerosum-strigosum-littorale)复合物的敲低转化体,用新构建的载体(pSA0104)应用粒子轰击,所述载体具有内源性 组成型启动子,其与对应于靶基因的反义链的DNA片段融合。 使用潮霉素抗性基因( aph7")选择潮霉素抗性菌落。 在第二次筛选后,通过PCR检查载体到基因组中的整合,并使用针对靶蛋白的特异性抗体通过蛋白质印迹评估敲低效应。
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