(参考ppt课件)自噬检测方法.ppt

自噬检测方法,1,2,3,常用细胞器标记物,4,5,6,自噬体检测：1.透射电镜2.免疫荧光观察LC3斑点,（P62辅助说明）；3.WB检测LC3+P62,并增加自噬降解抑制剂处理组。实验顺序：3 1,7,自噬流评价：1.双标LC3,红色斑点/黄色斑点;2.LC3与lamp1的共定位；3.DQ-BSA;4.稳转GFP-LC3细胞中检测游离GFP,8,Fusion of autophagosomes with endosomes/lysosomes is mediated by the class C VPS complex, Rab GTPase and the SNARE complex.,Cai, H., Reinisch, K. & Ferro-Novick, S. Coats, tethers, Rabs, and SNAREs work together to mediate the intracellular destination of a transport vesicle. Dev. Cell 12,671682 (2007). Itakura, E., Kishi-Itakura, C. & Mizushima, N. The hairpin-type tail-anchored SNARE syntaxin 17 targets to autophagosomes for fusion with endosomes/lysosomes. Cell 151, 12561269 (2012).Takts, S. et al. Autophagosomal Syntaxin17-dependent lysosomal degradation maintains neuronal function in Drosophila. J. Cell Biol. 201, 531539 (2013),机制研究：CO-IP比较各组处理后stx17结合的SNAP29和vamp8,9,