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HBSS, calcium, magnesium, no phenol red

HBSS,钙,镁,无酚红

公司名称: Thermo Fisher Scientific
产品编号: 14025126
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Fluorescence Microscopy Analysis of Drug Effect on Autophagosome Formation
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2014-04-05
[Abstract]  The autophagy protein, LC3 represents a reliable characteristic marker for autophagosomal structures. The initial LC3 is processed by the cysteine protease autophagy-related gene 4 (Atg4) at its C terminus in order to create LC3-I generally localized in the cytoplasm. Afterwards LC3-I is conjugated with phosphatidylethanolamine (PE) to become LC3-PE or LC3-II predominantly localised on the autophagosomal membranes (outer and inner). Autolysosomal content of LC3-II is very low as upon autophago/lysosomal fusion it is either cleaved off from the outer membrane by Atg4 or degraded together with ... [摘要]  自噬蛋白,LC3代表自噬体结构的可靠特征标记。最初的LC3由半胱氨酸蛋白酶自噬相关基因4(Atg4)在其C末端处理,以产生通常位于细胞质中的LC3-I。之后LC3-I与磷脂酰乙醇胺(PE)缀合以变成主要定位在自噬体膜(外部和内部)上的LC3-PE或LC3-II。 LC3-II的自溶酶体内含物非常低,因为在自噬/溶酶体融合时,其被Atg4从外膜切割或与内膜一起通过溶酶体活性降解。因此,GFP-LC3和mCherry-GFP-LC3可能通过常规或共聚焦荧光显微镜(FM)可视化。在这种情况下,mCherry-GFP-LC3或GFP-LC3细胞质池被可视化为均匀分散的信号,并且含有mCherry-GFP-LC3-II或GFP-LC3-II的自噬体被检测为点状形成。斑点数可以用作自噬体丰度的标志物。一般来说,我们建议计数每个细胞的GFP-LC3斑点的平均数。

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