Expansion Microscopy
By onExpansion microscopy is a technique to visualize biological structures with higher spatial resolution than traditional microscopy methods.
Microscopy-based GSH bead protein-protein interaction assay
By onProtocol describing the Microscopy-based GSH bead protein-protein interaction assay
Unconventional Initiation of PINK1/Parkin Mitophagy by Optineurin
By onCargo sequestration is a fundamental step of selective autophagy in which cells generate a double membrane structure termed an autophagosome on the surface of cargoes. The authors uncover an unconventional path of PINK1/Parkin mitophagy initiation.
circRNA dataset
By onNew data set on circRNA in laser-captured neuron samples from 190 human brains of healthy controls, prodromal PD, and clinical PD (Dong et al., Nature Communications, in press). The RNA-seq raw FASTQ data and normalized expression matrix of all circRNAs in this study have been deposited in GEO under accession number GSE218203.
circRNA Custom Code
By onCustom code associated with (Dong et al., Nature Communications, in press) is publicly available at https://github.com/sterding/circRNA.
Mutations in Parkinsonism-linked endocytic proteins synaptojanin1 and auxilin have synergistic effects on dopaminergic axonal pathology
By onParkinson's disease (PD) is a neurodegenerative disorder characterized by defective dopaminergic (DAergic) input to the striatum. Mutations in two genes encoding synaptically enriched clathrin-uncoating factors, synaptojanin 1 (SJ1) and auxilin, have been implicated in atypical Parkinsonism. SJ1 knock-in (SJ1-KIRQ) mice carrying a disease-linked mutation display neurological manifestations reminiscent of Parkinsonism. Here we report that auxilin knockout (Aux-KO) mice display dystrophic changes of a subset of nigrostriatal DAergic terminals similar to those of SJ1-KIRQ mice. Furthermore, Aux-KO/SJ1-KIRQ double mutant mice have shorter lifespan and more severe synaptic defects than single mutant mice. These include increase in dystrophic striatal nerve terminals positive for DAergic markers and for the PD risk protein SV2C, as well as adaptive changes in striatal interneurons. The synergistic effect of the two mutations demonstrates a special lability of DAergic neurons to defects in clathrin uncoating, with implications for PD pathogenesis in at least some forms of this condition.
pBPK1520-SNCA-A53T-ng
By onIt can be used to introduce SNCA-A53T mutation using prime editing, PE3 approach.
Live-cell imaging for synaptic vesicle precursors in human iNeuron axons
By onThe authors describe procedure and equipment used for live-imaging of synaptic vesicle precursors.
Global and APEX proteomics studying Golgi-phagy
By onDataset collection includes experiments on HEK, HeLa, iNeurons, and H9 ES cells under nutrient stress. Various proteomics analyses were conducted with different protein factors like FIP200, ATG7, LIR binding sites, YIPF4, YIPF3, and CALCOCO1.
Cell lysis and immunoblotting for protein and phospho-protein quantification
By onThe authors describe the procedure by which human iPSC-derived neurons or mouse embryonic fibroblasts (MEFs) were lysed and probed for levels of proteins of interest using Western blot.
Cellular lipid uptake with flow cytometry readout
By onCellular lipid uptake with flow cytometry readout
GST Bead pulldown Assay
By onGST Pulldown Assay for recruitment of bait proteins to GST labeled prey proteins. Prey proteins can be purified or from lysate.
Human neuroblastoma cell line SH-SY5Y culturing
By onThis is the brief protocol for seeding and culturing human neuroblastoma SH-SY5Y cells.
Endosomal and lysosomal immunoprecipitation for proteomics, lipidomics, and TEM
By onHere, authors describe an approach for purification of early/sorting endosomes, providing a means by which to examine early aspects of the endolysosomal system (Endo-IP) and to combine this with lysosome purification using Lyso-IP.
Live-cell imaging
By onLive-cell imaging is a technique to visualize dynamic cellular processes in living biological samples.
Tissue handling for mice, marmoset, and rhesus macaques injected with AAV
By onThis protocol outlines all procedures for handling of tissue of animal subjects treated with AAV.
