Native gel lipid binding assay
By savannah onThis protocol details how to perform a native gel lipid binding assay to evaluate the number of lipids bound per molecule of 3xFLAG-SHIP164Δ901-1099.
Molecular cloning of SHIP164 plasmids for expression in mammalian cells
By savannah onThis protocol is to help with the molecular cloning of SHIP164 and the sequences of other proteins.
Lipid transfer assay
By savannah onThis protocol details how to perform a lipid transfer assay with 3xFLAG-SHIP164Δ901-1099.
iPSC differentiation into Microglia
By savannah onThis protocol describes iPSC differentiation into microglia.
iPSC differentiation into Macrophages
By savannah onThis protocol describes iPSC differentiation into macrophages.
Immunofluorescence of macrophages and microglia
By savannah onThis protocol describes the fixation and staining of cultured cells in paraformaldehyde solution.
Immunoblotting of macrophages and microglia
By savannah onThis protocol describes the preparation from cell lysate from cultured cells and immunoblotting procedure.
Generation of knock-in and knock-out CRISPR/Cas9 editing in mammalian cells
By savannah onThis protocol is to help with generation of knock-in and knock-out CRISPR/Cas9 editing in mammalian cells.
Erythroid differentiation dependent interaction of VPS13A with XK at the plasma membrane of K562 cells
By savannah onPublished: ER-PM contacts positive for VPS13A were seldomly observed in undifferentiated K562 cells, despite the presence of XK in these cells at concentrations similar to those observed after differentiation. Findings reveal the interaction of VPS13A with XK at ER-PM contacts requires a permissive state which depends upon cell type and/or functional state of the cell. View original preprint.
Ectopic receptor & endosome size and count image analysis and quantification
By savannah onThis protocol is to help with the imaging analysis of ectopic receptor localization and endosome size.
DQ-BSA assay
By savannah onThis protocol describes the DQ BSA assay for measuring the lysosomal proteolytic activity.
Cryo-EM sample preparation
By savannah onThis protocol details how to prepare cryo-EM samples of 3xFLAG-SHIP164Δ901-1099.
Cell culture, transfection, immunocytochemistry, and imaging
By savannah onThis protocol is to help with the maintenance, transfection, immunocytochemistry, and imaging of adherent mammalian cells.
Bone Marrow Derived Macrophage (BMDM) differentiation and maintenance
By savannah onThis protocol describes the isolation of bone marrow derived macrophages.
3D FIBSEM CLEM
By savannah onThis protocol details the procedure of Correlative Light Microscopy and Electron Microscopy (CLEM) with 3D Focus Ion Beam Scanning Electron Microscopy (FIBSEM) technique in Zeiss Crossbeam 550 FIBSEM system.
Inhibition of striatal dopamine release by the L-type calcium channel inhibitor isradipine co-varies with risk factors for Parkinson’s
By savannah onPublished: This data show that LTCC function in DA axons, and isradipine effect, are locally governed and suggest they vary in a manner that in turn might impact on, or reflect, the cellular stress that leads to parkinsonian degeneration. View original preprint.
Synaptic location is a determinant of the detrimental effects of α-synuclein pathology to glutamatergic transmission in the basolateral amygdala
By savannah onαSyn expression is restricted in a subset of glutamatergic synapses in BLA and its aggregation decreases cortico-BLA transmission through both gained toxicity and loss of normal function. These results might be relevant to the reduced cortical control of amygdala function that has been associated with psychiatric deficits in PD.
Vibrational stabilization of cluster synchronization in oscillator networks
By savannah onPreprint: Deviations from normal cluster synchronization patterns are closely associated with various malfunctions, such as neurological disorders in the brain. Here, the authors employ an open-loop control strategy, vibrational control, which does not require any state measurements. Additionally, they conduct numerical experiments to demonstrate their theoretical findings.
Sample preparation for TMT-based total and phospho-proteomic analysis of cells and tissues
By savannah onMass spectrometry-based proteomics and phosphoproteomics are highly sensitive and un-biased techniques to study the proteome and phosphoproteome at a global scale. Using our protocols, we routinely identify and quantify >35,000 phosphosites and ~10,000 protein groups.