自噬检测方法

自噬检测方法常用细胞器标记物自噬体检测：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：31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GFPFusionofautophagosomeswithendosomes/lysosomesismediatedbytheclassCVPScomplex,RabGTPaseandtheSNAREcomplex.Cai,H.,Reinisch,K.&Ferro-Novick,S.Coats,tethers,Rabs,andSNAREsworktogethertomediatetheintracellulardestinationofatransportvesicle.Dev.Cell12,671–682(2007).Itakura,E.,Kishi-Itakura,C.&Mizushima,N.Thehairpin-typetail-anchoredSNAREsyntaxin17targetstoautophagosomesforfusionwithendosomes/lysosomes.Cell151,1256–1269(2012).Takáts,S.etal.AutophagosomalSyntaxin17-dependentlysosomaldegradationmaintainsneuronalfunctioninDrosophila.J.CellBiol.201,531–539(2013)机制研究：CO-IP比较各组处理后stx17结合的SNAP29和vamp8