Quick guide to use paceTOMO for cryo-ET data collection from Titan Krios
By onThis quick guide provides key minimal steps for preparing the Titan/SerialEM for the tomogram data collection on lamella or in vitro specimens with a K3 camera. paceTOMO routine is also included for a typical tomogram data collection session. Please note that this is not an exhaustive guide, but summarises the order of key steps.
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).
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.
Neural differentiation on EM grids – iNeurons sample preparation for cryo-ET and CLEM
By onProtocol for direct differentiation of AAVS1-TRE3G-NGN2 iPSCs and hESCs to iNeurons on EM grids: coat grids, seed iNeurons, transduce with fluorescent proteins, then plunge freeze for cryo-ET and cryo-CLEM.
Expression and purification protocol of GST-mCh-FYVE
By onThis protocol details the expression and purification of GST-mCh-FYVE.
Global proteomic analysis of WT and GRN-/- HeLa cells
By onGlobal proteomic analysis of WT and GRN-/- HeLa cells https://www.nature.com/articles/s41467-022-33500-9
Generation of stable cell lines via retroviral or lentiviral transduction
By onThis protocol details how to generate stable cell lines using a retrovirus system
Quantifying Acetylcholinesterase activity in striatal brain tissue with DTNB assay
By onThis protocol describes the steps to measure acetylcholinesterase (AChE) activity in mouse striatal brain tissue, including how to store tissue samples, extract AChE and obtain Michaelis-Menten like plots to measure the rate of AChE activity.
Sectioning of Mouse Brain by Microtome
By onThis protocol describes how to use the microtome to prepare and slice mouse brain sections for Immunohistochemistry.
mCherry-YIPF4 Immunoprecipitation V2
By onThe protocol outlines how to immunoprecipitate mCherry-YIPF4 for further analysis.
Immunoblotting of macrophages and microglia
By onThis protocol describes the preparation from cell lysate from cultured cells and immunoblotting procedure.
Mitochondrial complex activity assays
By onMitochondria complex activity assays measure the activity levels of the different complexes of the mitochondrial electron transport chain (ETC).
Reconstitution of cargo-induced LC3 lipidation in mammalian selective autophagy
By onCode for analysis of multicolor fluorescence images to quantify fluorescence intensity at the perimeter of giant unilamellar vesicles (GUVs).
Cryo-EM sample preparation
By onThis protocol details how to prepare cryo-EM samples of 3xFLAG-SHIP164Δ901-1099
Midbrain viral injections for striatal fiber photometry in mice
By onThis protocol describes how to: - Inject AAV viruses into the midbrain (SNc: substantia nigra pars compacta, or VTA: ventral tegmental area) of mice - Train mice for head-fixed running on a cylindrical treadmill, and to receive rewards and air puffs while head-fixed - Implant head plates on mice for head-fixation during behavior (optional) While this protocol is focused on injections of AAV viruses into the midbrain, it can be easily modified to inject into other brain regions or other viruses.
Structural basis for ATG9A recruitment to the ULK1 complex in mitophagy initiation
By onHere, the authors examine the structural interaction between ATG9A and components of the ULK1 complex to better understand the process of the PINK1- and Parkin- dependent mitophagy pathway implicated in Parkinson's disease.
