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.
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.
Immunostaining of iPSC-derived neurons for quantification of synaptic proteins
By onHere, the authors fix, permeabilize, and stain human iPSC-derived neurons for the purpose of observing and quantifying somal proteins of interest.
Structure of human ULK1 complex core (2:1:1 stoichiometry)
By onStructure of human ULK1 complex core (2:1:1 stoichiometry)
Mitoguardin-2–mediated lipid transfer preserves mitochondrial morphology and lipid droplet formation
By onLipid transport proteins at membrane contacts, where organelles are closely apposed, are critical in redistributing lipids from the endoplasmic reticulum (ER), where they are made, to other cellular membranes. Such protein-mediated transfer is especially important for maintaining organelles disconnected from secretory pathways, like mitochondria. We identify mitoguardin-2, a mitochondrial protein at contacts with the ER and/or lipid droplets (LDs), as a lipid transporter. An x-ray structure shows that the C-terminal domain of mitoguardin-2 has a hydrophobic cavity that binds lipids. Mass spectrometry analysis reveals that both glycerophospholipids and free-fatty acids co-purify with mitoguardin-2 from cells, and that each mitoguardin-2 can accommodate up to two lipids. Mitoguardin-2 transfers glycerophospholipids between membranes in vitro, and this transport ability is required for roles both in mitochondrial and LD biology. While it is not established that protein-mediated transfer at contacts plays a role in LD metabolism, our findings raise the possibility that mitoguardin-2 functions in transporting fatty acids and glycerophospholipids at mitochondria-LD contacts.
Parkinson’s disease and cancer: a systematic review and meta-analysis of over 17 million participants
By onThe authors examined risk association between Parkinson’s disease and cancer using data from 63 publications. With the exception of melanoma, the authors found that the risk association of Parkinson’s disease and cancer was inversely related.
LRRK2 Immunofluorescent staining
By onProtocol for immunofluorescent staining for LRRK2 in cultured cells using the MJFF2 (c41-2) antibody.
Towards a phenome-wide view of Parkinson’s disease
By onMany studies have examined the relation between PD and environmental variables serially --- one candidate association at a time. In the real world however, both environmental exposures and patients are much more complex, including correlated environmental exposures, polypharmacy, and complex comorbidities. Here we begin to characterize a holistic view of environmental, health, and pharmacological traits linked to patients with PD.
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.
Image processing of full-length monomeric LRRK2
By onThis protocol assumes that 2000 to 4000 movies of full-length LRRK2 embedded on vitreous ice are collected with an electron microscope equipped with a direct detector. It focuses primarily on the monomeric population of LRRK2 and has been leading to 3-4 Å resolution structures.
H9 ES AAVS1-NGN2 FAM134C/A/B-/-;TEX264-/-; CCPG1-/-; PiggyBac-Keima-RAMP4
By onES cells were used to create iNeurons lacking ER-phagy receptor genes FAM134C, A, B, TEX264, CCPG1. They expressed Keima-RAMP4 ER-phagy flux reporter. CRISPR/Cas9 was used to introduce NEUROG2 construct in AAVS1 safe harbor locus.
Isolation and Processing of Embryonic and Postnatal Brains
By onThis protocol describes: -Harvesting embryos from females from embryonic day 10 to embryonic day 18 -Brain isolation of embryonic (e10-e18) and postnatal brains (p0-p30) -Processing and Freezing of embryonic and postnatal brains
Structure of PPM1H phosphatase with manganese ions at the active site
By onStructure of PPM1H phosphatase with manganese ions at the active site (as reported in 10.15252/embr.202152675) deposited in the Protein Data Bank with code 7N0Z.
Cell culture on EM grids and fluorescence microscopy imaging
By onThis protocol describes the general procedure of seeding mammalian cells on EM grids and confocal fluorescence microscopy imaging of grids for subsequent cryo-tomography experiments, performed.
Hydrop enables droplet-based single-cell ATAC-seq and single-cell RNA-seq using dissolvable hydrogel beads
By onHyDrop protocol enables single-cell sequencing for RNA and chromatin accessibility. Generated over 7996 single-cell profiles for ATAC-seq and 9508 for RNA-seq in mouse cortex. Protocol validated on low-input samples from Drosophila brain.
Structure of human ULK1 complex core (2:2:2 stoichiometry) in the PI3KC3-C1 mixture
By onStructure of human ULK1 complex core (2:2:2 stoichiometry) in the PI3KC3-C1 mixture (Method: ELECTRON MICROSCOPY, Resolution: 5.85 Å, Aggregation State: PARTICLE, Reconstruction Method: SINGLE PARTICLE).