Endo-IP and Lyso-IP Toolkit for Endolysosomal Profiling of Human Induced Neurons

We developed a toolkit for capturing endosomes (Endo-IP) and lysosomes (Lyso-IP) in human stem cell(hESC)-derived induced neurons (iNeurons). We target a sub-population of early endosomes marked by endogenously tagged EEA1 (Early Endosome Antigen 1) protein, which represents one of the earliest populations formed after endocytic vesicle uncoating. We demonstrated its utility by revealing the endolysosomal protein landscapes for cortical-like iNeurons and stem cells. This allowed us to globally profile endocytic cargo, identifying hundreds of transmembrane proteins.