Generation of stable cell lines using viral infection
By onProtocol for generation and precipitation of retrovirus, and infection of HeLa cells to generate stable cell lines.
Circular RNA in the human brain are tailored to neuron identity and neuropsychiatric disease
By onLittle is known about circular RNAs (circRNAs) in specific brain cells and human neuropsychiatric disease. Here, we systematically identify over 11,039 circRNAs expressed in vulnerable dopamine and pyramidal neurons laser-captured from 190 human brains and non-neuronal cells using ultra-deep, total RNA sequencing. 1526 and 3308 circRNAs are custom-tailored to the cell identity of dopamine and pyramidal neurons and enriched in synapse pathways. 29% of Parkinson's and 12% of Alzheimer's disease-associated genes produced validated circRNAs. circDNAJC6, which is transcribed from a juvenile-onset Parkinson's gene, is already dysregulated during prodromal, onset stages of common Parkinson's disease neuropathology. Globally, addiction-associated genes preferentially produce circRNAs in dopamine neurons, autism-associated genes in pyramidal neurons, and cancers in non-neuronal cells. This study shows that circular RNAs in the human brain are tailored to neuron identity and implicate circRNA-regulated synaptic specialization in neuropsychiatric diseases.
Testing AAVrg-hSyn-mCherry in the Spinal Cord of Rhesus Macaques
By onViral Vector: Two rhesus macaque animals were tested for AAVrg-hSyn-mCherry in the spinal cord. The authors observed abundant mCherry-positive neurons in expected targets (i.e., motor cortical regions known to project to the spinal cord).
Testing AAVrg-hSyn-mCherry in the Putamen of Rhesus Macaques
By onTwo rhesus macaques were tested with AAVrg-hSyn-mCherry in the putamen, showing numerous mCherry-positive neurons in motor cortical regions projecting to the putamen.
A computational pipeline to quantify perinuclear lysosomes in fibroblasts using CellProfiler
By onHere we present a CellProfiler software pipeline to quantify the distribution of lysosomes in MEF cells. The lysosomes were stained using anti-LAMP1 antibody, and nuclei were labeled using DAPI. The images were acquired using a Zeiss laser scanning confocal microscope and were maximum intensity projected in FIJI.
Video tutorials for Curtain and Curtain-PTM
By onVideo tutorials to assist users of Curtain and Curtain-PTM proteomic data analysis web tools (described in ).
Calculating mitochondrial protein solubility
By onProtocol for generating solubility calculations for mitochondrial proteins from starting 'soluble' and 'insoluble' fraction mass spectrometry data, using MaxQuant and Perseus software pipelines.
Reverse-phase high pH fractionation (using Thermo Fisher, Cat# 84868)
By onThis protocol uses the Pierce™ High pH Reversed-Phase Peptide Fractionation Kit (Thermo Fisher, Cat# 84868)
Preparation of soluble and insoluble mitochondrial protein fractions for mass spectrometry analysis
By onProtocol describing preparation of soluble and insoluble mitochondrial protein fractions from HeLa cells for mass spectrometry analysis.
Preparation of soluble and insoluble mitochondrial protein fractions for immunoblotting
By onProtocol describing the preparation procedure of soluble and insoluble mitochondrial protein fractions from HeLa cells for immunoblotting.
Mitochondrial isolation from HeLa cells
By onMitochondrial isolation and quantification from HeLa cells.
Western blotting using the BioRad Criterion Blotter system
By onProtocol for performing an SDS-PAGE Western blot analysis using the BioRad Criterion Blotter system.
Preparing whole cell samples for immunoblot analysis
By onProtocol for preparation of HeLa cell lysates for immunoblot analysis.
Induction of starvation stress
By onProtocol describing the induction of starvation stress using EBSS in HeLa cells
Immunofluorescence Assay (IFA)
By onThis protocol details the procedure of immunofluorescence assay (IFA).
qRT-PCR sample preparation
By onProtocol for qRT-PCR sample preparation for analysis using a RotorGeneQ machine (Qiagen).
Analyzing oxygen consumption of isolated mitochondria using the Seahorse XFe96 analyzer
By onProtocol for analyzing oxygen consumption of isolated mitochondria using the Seahorse XFe96 analyzer.
Analyzing cellular ATP levels
By onProtocol for analysis of cellular ATP levels using the Promega Mitochondrial ToxGlo Kit.
Measuring mitophagy via FACS with mtKeima reporter
By onPreparation of samples for measuring mitophagy levels using mtKeima reporter by fluorescence activated cell sorting (FACS).
