Cell viability assessment
By Emma Sherrell onThis is the protocol for assessing cell viability using a MTS cell proliferation assay kit.
Evaluating GPNMB ACD mutants by Western Blotting and immunofluorescence
By Julia Julia Leonard onThis protocol describes the evaluation of cellular processing of GPNMB mutants by Western Blotting and Immunofluorescent imaging in a HEK293 overexpression model.
Mitochondrial antigen presentation in RAW macrophages
By Emma Sherrell onThis protocol details methods for 3-day Mitochondrial Antigen Presentation Assay in a murine macrophage cell line (RAW) that expresses a glycoprotein B (gB) of herpes simplex virus 1 (HSV1) targeted to the mitochondrial matrix (mito-gB). Watch an interview about this protocol with Ahmed Fahmy, PhD.
Loss of primary cilia and dopaminergic neuroprotection in pathogenic LRRK2-driven and idiopathic Parkinson’s disease
By Julia Julia Leonard onActivating LRRK2 mutations causes Parkinson's disease. Previously, work showed that cholinergic interneurons and astrocytes lose primary cilia in LRRK2 mutant mice. Here, the data strongly suggest that loss of cilia in specific striatal cell types decreases neuroprotection for dopamine neurons in mice and human Parkinson's disease.
Neural differentiation on EM grids – iNeurons sample preparation for cryo-ET and CLEM
By Emma Sherrell onA protocol for differentiating AAVS1-TRE3G-NGN2 hESCs to iNeurons directly on EM grids for cryo-ET and cryo-CLEM. The protocol includes: (1) EM grids coating with Matrigel or Poly-Ornithine and Laminin; (2) differentiation and seeding of iNeurons on EM grids; (3) lentiviral transduction for transient expression of fluorescently-labeled proteins; and (4) plunge freezing. Watch an interview about this protocol with Cristina Capitanio, MSc.
Primary data associated with the manuscript 10.1126/science.adi9926 (“Rab29-dependent asymmetrical activation of leucine-rich repeat kinase 2”)
By Julia Julia Leonard onPrimary data associated with the manuscript "Rab29-dependent asymmetrical activation of leucine-rich repeat kinase 2" (Hanwen Zhu, Francesca Tonelli, Martin Turk, Alan Prescott, Dario R. Alessi, Ji Sun). These include immunoblotting data and their quantitation, representative confocal microscopy images, and quantitation and statistical analysis of confocal microscopy images.
Peripheral MC1R activation modulates immune responses and confers neuroprotection in a mouse model of Parkinson’s disease
By Emma Sherrell onDataset corresponding to the article, "Peripheral MC1R activation modulates immune responses and confers neuroprotection in a mouse model of Parkinson's disease."
Nigrostriatal organotypic cultures to study neuromelanin accumulation in dopaminergic circuits
By Julia Julia Leonard onIn this protocol, the authors describe the preparation and maintenance of rat organotypic cultures from parasagittal brain slices.
FASTQ alignment, gene counts, and differential expression analysis and functional annotation of DEGs
By Julia Julia Leonard onFASTQ alignment, gene counts, and differential expression analysis and functional annotation of DEGs.
Patient PBMC differentiation to macrophages
By Julia Julia Leonard onPatient PBMC differentiation to macrophages.
pH-Rodo pathogens to monitor phagocytosis in pMacs
By Julia Julia Leonard onpH-Rodo pathogens to monitor phagocytosis in pMacs.
DQ-BSA and BMV109 microscopy
By Julia Julia Leonard onDQ-BSA and BMV109 microscopy for peritoneal macrophages.
Nigrostriatal Tau pathology in Parkinsonism and Parkinson’s disease
By Emma Sherrell onDataset corresponding to the preprint, "Is Tau the Initial Pathology in Dopaminergic Nigrostriatal Degeneration? Studies in Parkinsonism and Parkinson’s Disease."
Structural analysis of 20S CPs and assembly intermediates by electron cryo-microscopy
By Emma Sherrell onThis protocol details methods for structural determination by transmission electron cryo-microscopy of 20S CPs and assembly intermediates.
Flow cytometry for ex vivo stimulated pMacs
By Julia Julia Leonard onFlow cytometry for ex vivo stimulated pMacs.
Cloning, protein expression, and purification of 20S CPs and assembly intermediates
By Emma Sherrell onThis protocol details methods for cloning, expression, and purification of 20S CPs, and assembly intermediates for biochemical and structural analysis.
Open-source platform for kinematic analysis of mouse forelimb movement
By Emma Sherrell onAn open-source behavioral platform and software solution for studying fine motor skills in mice performing reach-to-grasp tasks. The behavioral platform uses readily available and 3D-printed components and was designed to be affordable and universally reproducible.
3D bioprinting of human neural tissues with functional connectivity
By Emma Sherrell onProbing how human neural networks operate is hindered by the lack of reliable human neural tissues amenable to the dynamic functional assessment of neural circuits. Team Scherzer developed a 3D bioprinting platform to assemble tissues with defined human neural cell types in a desired dimension using a commercial bioprinter.