Culturing, transfection and NBD-labeled lipid uptake assay in HeLa-CDC50A-OE cells
By onAssay for NBD-labeled lipid uptake in HeLa-CDC50A-OE cells includes seeding, transfection, and flow cytometry to measure flippase construct activity, enabling quantitative assessment of lipid uptake.
Flow cytometry data from the manuscript “Fluorescence-Gated Flow Cytometry Approach for Measuring Lipid Flippase Activity in Mamma-lian Cells”
By onSummary: The manuscript presents a fluorescence-gated flow cytometry method to measure lipid flippase activity in mammalian cells.
Spermidine suppresses glial inflammation and parkinsonian abnormalities in ATP13A2 deficiency
By onPathogenic variants in ATP13A2 cause Kufor-Rakeb syndrome & early-onset parkinsonism. Spermidine supplementation rescues symptoms by counteracting microglia dysfunction & improving neuronal integrity.
NERINE reveals rare variant associations in gene networks across phenotypes and implicates an SNCA-PRL-LRRK2 subnetwork in Parkinson’s disease
By onNERINE is a rare variant association test that links rare genetic variation with gene network topology to improve discovery power in complex diseases. NERINE implicates an α-synuclein/prolactin stress-response in Parkinson’s disease.
all protocols related to “Spermidine suppresses glial inflammation and parkinsonian abnormalities in ATP13A2 deficiency”
By onAll protocols related to "Spermidine suppresses glial inflammation and parkinsonian abnormalities in ATP13A2 deficiency"
Regulation of the proteasome 20S core particle by the Parkinsonism-associated Proteins FBXO7 and PI31
By onGenetic forms of PD are associated with variants in FBXO7 and PI31. Combining biochemical and structural approaches, we describe the proteasome interaction of these proteins and map disease variants to disruption of proteasome interaction.
Fluorescence-Gated Flow Cytometry Approach for Measuring Lipid Flippase Activity in Mammalian Cells
By onP4-ATPase lipid flippases create lipid asymmetry in cells. A new strategy using NBD-lipid uptake assays improves sensitivity and analysis of ATP11C function. This method enhances studying regulatory interactions in mammalian cells.
pCHMWS-hATP13A4_A356V-IRES-Puro
By ontransfer plasmid for lentiviral vector production expressing Hs ATP13A4 A356V mutant
pCHMWS-hATP13A4_A356V-Twinstrep-Flag-IRES-Puro
By ontransfer plasmid for lentiviral vector production expressing Hs ATP13A4 A356V mutant with Twinstrep-Flag tag
AAV-gfaABC1D-Lamp1-TurboID-HA
By onUsing the gfaABC1D promoter expressing Lamp1-TurboID fusion protein
pCHMWS-FLUC-IRES-Puro
By ontransfer plasmid for lentiviral vector production expressing firefly luciferase (FLUC)
AAV-gfaABC1D-cytoTurboID-HA
By onUsing the gfaABC1D promoter expressing cytoplasmic TurboID with C-terminal HA tag
Postnatal astrocyte labeling by electroporation
By onHere, we provide a step-by-step guide to perform Postnatal Astrocyte Labeling by Electroporation (PALE), detailing the preparation, electroporation procedure, and post-electroporation analysis.
in vivo TurboID
By onProtocol uses iBioID with Lamp1-TurboID in astrocytes to study endo-/lysosomal protein interactions in mice. AAV delivery allows selective biotinylation for proteome mapping, offering insights into astrocyte signaling in vivo.
Immunocytochemistry – subcellular localization of ATP13A4
By onThis protocol describes an optimized method for visualizing the subcellular localization of over-expressed ATP13A4 in HeLa cells using immunocytochemistry.
IHC staining (DAB) of Iba1 and Gfap
By onThis protocol details the IHC staining (DAB) of Iba1 and Gfap.
Protein quantification by SDS-PAGE and Coomassie staining
By onThis protocol provides a step-by-step guide for quantifying protein concentration by SDS-PAGE and Coomassie staining.
Immunoblotting protocol
By onThis protocol outlines a step-by-step method for performing western blot analysis, including protein separation by gel electrophoresis, transfer to membranes, antibody incubation, and detection.
Expression and purification of ATP13A4 from Saccharomyces cerevisiae
By onThis protocol describes the expression and purification of human ATP13A4 with a C-terminal Twin-Strep tag from Saccharomyces cerevisiae using a CuSO₄-inducible system.
