Raw tomogram of a TauRD-YFP aggregate in a TauRD-YFP expressing HEK293T cells and primary mouse neurons.
By onRaw tomogram of a TauRD-YFP aggregate in a TauRD-YFP expressing Hek293T cells. Accession numbers: HEK293T: https://www.ebi.ac.uk/emdb/EMD-13739 Primary mouse neurons: https://www.ebi.ac.uk/emdb/EMD-13740
Plunge freezing
By onThis protocol describes the procedure of plunge freezing for subsequent cryo-electron tomography.
Immunofluorescence staining for postmortem mouse brain tissue
By onThis is a basic protocol for staining mouse brain tissues using immunofluorescence and immunohistochemistry techniques.
Single-molecule Immunofluorescence Tissue Staining Protocol for Oligomer Imaging V.3
By onThis protocol details background fluorescence quenching and immunofluorescence staining of human brain tissue for oligomer imaging.
Cell culture, transfection and imaging
By onThis protocol details the general preparation of cells for imaging and also for imaging experiments involving cellular hypotonic shock and cytosolic Ca2+ changes as they were performed in https://doi.org/10.1083/jcb.202010004.
Immunocytochemical analysis
By onAssociated with preprint: https://www.biorxiv.org/content/10.1101/2022.11.02.514817v1
Alpha-synuclein immunochemistry on STC-1 cells using DAB
By onThis protocol describes how to visualise alpha-synuclein in STC-1 cells by DAB immunohistochemistry. It also works for other antibodies (e.g. 5-HT, CCK, GLP).
Data set of the manuscript “Neuronal hyperactivity-induced oxidant stress promotes in vivo a-synuclein brain spreading”
By onData set of the manuscript "Neuronal hyperactivity-induced oxidant stress promotes in vivo a-synuclein brain spreading."
Regulatory imbalance between LRRK2 kinase, PPM1H phosphatase, and Arf6 GTPase disrupts the axonal transport of autophagosomes
By onThe authors found that LRRK2-hyperphosphorylated RABs disrupt the axonal transport of autophagosomes by perturbing the coordinated regulation of cytoplasmic dynein and kinesin motors.
Images of adeno-associated viral vectors for functional intravenous gene transfer throughout the non-human primate brain
By onDataset of images supporting the publication: Chuapoco, M.R., Flytzanis, N.C., Goeden, N. et al. Adeno-associated viral vectors for functional intravenous gene transfer throughout the non-human primate brain. Nat. Nanotechnol. (2023). https://doi.org/10.1038/s41565-023-01419-x Publication Abstract: Crossing the blood-brain-barrier in non-human primates (NHPs) is a major obstacle for gene delivery to the brain. Adeno-associated viruses (AAVs) promise robust gene delivery to the brain through non-invasive, intravenous delivery. However, unlike in rodents, few neurotropic AAVs efficiently cross the blood-brain barrier in non-human primates. Here, we report on AAV.CAP-Mac, an engineered variant identified by screening in adult marmosets and new-born macaques which has improved delivery efficiency in the brains of multiple NHP species: marmoset, rhesus macaque, and green monkey. CAP-Mac is neuron-biased in infant Old-World primates, exhibits broad tropism in adult rhesus macaques, and is vasculature-biased in adult marmosets. We demonstrate applications of a single, intravenous dose of CAP-Mac to deliver functional GCaMP for ex vivo calcium imaging across multiple brain areas, or a cocktail of fluorescent reporters for Brainbow-like labeling throughout the macaque brain, circumventing the need for germline manipulations in Old World primates. As such, CAP-Mac is shown to have potential for non-invasive systemic gene transfer in the brains of NHPs.
Reconstitution of cargo-induced LC3 lipidation in mammalian selective autophagy
By onGUV quantification
ENS Quantification
By onLabelled enteric neurons in living mice with recombinant adeno-associated viruses (rAAVs) expressing fluorescent proteins, and used tissue clearing techniques to enhance visualization of intact GI tissue.
Expansion microscopy with R1441C LRRK2 MEF cells: visualization of Myc-RILPL1 and TMEM55B
By onExpansion microscopy is a super-resolution imaging technique that increases the physical distance between fluorophores from fixed cells on coverslips. Here, the authors use expansion techniques to confocal image TMEM55B and RILPL1.
Preparation and imaging of enriched Golgi from GolgiTAG-IP using Transmission Electron Microscopy
By onA protocol where Golgi, isolated from cells by GolgiTAG immunoprecipitation (IP) can be prepared and imaged using TEM. This protocol can also be used to image any organelles isolated using various organelle-IP protocols that are available.
immunofluorescent staining with anti-GFP and anti-CD63 antibodies
By onimmunofluorescent staining with anti-GFP and anti-CD63 antibodies
Fixing hippo neurons to assess endogenous NEMO during oxidative stress
By onProtocol describing the procedure for fixing Hippocampal rat neurons to assess endogenous NEMO during oxidative stress.
Preparation of LRRK2 RCKW cryo-EM grids
By onThis is Leschziner's Lab updated protocol for making cryo-EM grids for LRRK2 RCKW. This protocol, when using lower protein concentration, results in better monomer and dimer formation than the old protocol.
Fixation of HeLa-M cells expressing Halo and SNAP fusion proteins conjugated to ligands
By onProtocol describing fixation of HeLa-M cells expressing Halo and SNAP fusion proteins conjugated to ligands.
Immunofluorescence Assay (IFA)
By onThis protocol details the procedure of immunofluorescence assay (IFA).
