Microscopy-based GSH bead protein-protein interaction assay
By onProtocol describing the Microscopy-based GSH bead protein-protein interaction assay
Cellular lipid uptake with flow cytometry readout
By onCellular lipid uptake with flow cytometry readout
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.
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.
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.
Proteomics workflow for APP/Aβ TOMAHAQ analysis in endosomal and lysosomal fractions
By onProtocol for analyzing Ab "half-tryptic" peptides from organelles using TOMAHAQ and Tomahto on a Thermo orbitrap instrument for detecting APP processing to Ab amyloid peptide.
Mitomycin C inactivation of mouse embryonic fibroblasts (MEFs) for hPSC cultures
By onThis protocol describes the process of using Mitomycin C to inactivate mouse embryonic fibroblasts (MEFs), which can then be used as feeder cells for human pluripotent stem cell (hPSC) culture.
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.
Live-cell imaging
By onLive-cell imaging is a technique to visualize dynamic cellular processes in living biological samples.
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.
Human neuroblastoma cell line SH-SY5Y culturing
By onThis is the brief protocol for seeding and culturing human neuroblastoma SH-SY5Y cells.
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.
In vitro assembling of RNP for nucleofection of hPSCs
By onThis protocol describes the procedure for the in vitro assembly of ribonucleoprotein (RNP) which can be delivered into human pluripotent stem cells (hPSCs) using nucleofection.
