'Sorting of RNAs to specific subcellular loci occurs in diverse settings from fly oocytes to mammalian neurons. Using the membrane-permeable nucleic acid stain SYTO 14, we directly visualized the translocation of endogenous RNA in living cells. Labeled RNA was distributed nonrandomly as discrete granules in neuronal processes. The labeled granules ... More
Nucleic acid dyes for detection of apoptosis in live cells.
AuthorsFrey T
JournalCytometry
PubMed ID8582249
'Apoptotic thymocytes were found to be much dimmer than normal thymocytes when stained with several nucleic acid dyes. These dyes provide a quick and simple assay for apoptosis which works for live cells and does not require a UV laser. The collection of dyes giving this staining pattern includes reagents ... More
A rapid sample preparation technique for flow cytometric analysis of immunofluorescence allowing absolute enumeration of cell subpopulations.
AuthorsTerstappen LW, Meiners H, Loken MR
JournalJ Immunol Methods
PubMed ID2477460
'A simple and rapid method was developed for immunofluorescence measurements of cells by flow cytometry which does not require washing procedures, permitting absolute enumeration of cell subpopulations. Peripheral blood cells were labeled with fluorescein and phycoerythrin conjugated monoclonal antibodies and the nucleic acid stain LDS-751. Distilled water was added following ... More
Neocentromere-mediated chromosome movement in maize.
AuthorsYu HG, Hiatt EN, Chan A, Sweeney M, Dawe RK
JournalJ Cell Biol
PubMed ID9362502
'Neocentromere activity is a classic example of nonkinetochore chromosome movement. In maize, neocentromeres are induced by a gene or genes on Abnormal chromosome 10 (Ab10) which causes heterochromatic knobs to move poleward at meiotic anaphase. Here we describe experiments that test how neocentromere activity affects the function of linked centromere/kinetochores ... More
Disrupted-in-schizophrenia 1 regulates transport of ITPR1 mRNA for synaptic plasticity.
AuthorsTsuboi D, Kuroda K, Tanaka M, Namba T, Iizuka Y, Taya S, Shinoda T, Hikita T, Muraoka S, Iizuka M, Nimura A, Mizoguchi A, Shiina N, Sokabe M, Okano H, Mikoshiba K, Kaibuchi K
Journal
PubMed ID25821909
'Disrupted-in-schizophrenia 1 (DISC1) is a susceptibility gene for major psychiatric disorders, including schizophrenia. DISC1 has been implicated in neurodevelopment in relation to scaffolding signal complexes. Here we used proteomic analysis to screen for DISC1 interactors and identified several RNA-binding proteins, such as hematopoietic zinc finger (HZF), that act as components ... More
Deep ultraviolet mapping of intracellular protein and nucleic acid in femtograms per pixel.
'By using imaging spectrophotometry with paired images in the 200- to 280-nm wavelength range, we have directly mapped intracellular nucleic acid and protein distributions across a population of Chinese hamster ovary (CHO-K1) cells. A broadband 100× objective with a numerical aperture of 1.2 NA (glycerin immersion) and a novel laser-induced-plasma ... More
Detection of apoptosis in live cells by MitoTracker red CMXRos and SYTO dye flow cytometry.
AuthorsPoot M, Gibson LL, Singer VL
JournalCytometry
PubMed ID9098628
'We characterized the ability of six SYTO nucleic acid stains and a mitochondrial stain to resolve by flow cytometry camptothecin-induced apoptotic and non-apoptotic cells. Staining live human lymphoid B-cells showed such resolution with SYTO 11, 12, 13, 14, and 16 dyes. H9, HL-60, and Jurkat cells did not show resolution ... More
Microtubule-dependent recruitment of Staufen-green fluorescent protein into large RNA-containing granules and subsequent dendritic transport in living hippocampal neurons.
AuthorsKöhrmann M, Luo M, Kaether C, DesGroseillers L, Dotti CG, Kiebler MA
JournalMol Biol Cell
PubMed ID10473638
'Dendritic mRNA transport and local translation at individual potentiated synapses may represent an elegant way to form synaptic memory. Recently, we characterized Staufen, a double-stranded RNA-binding protein, in rat hippocampal neurons and showed its presence in large RNA-containing granules, which colocalize with microtubules in dendrites. In this paper, we transiently ... More
Neurotrophin-3 signals redistribute RNA in neurons.
AuthorsKnowles RB, Kosik KS
JournalProc Natl Acad Sci U S A
PubMed ID9405694
The translocation of specific mRNAs to dendrites and their potential for locally regulated translation are likely to serve as an effector in neuronal plasticity. Whether translation in dendrites is regulated by delivery of the RNA to sites of plasticity or a stationary pool of localized RNA undergoes enhanced translational efficiency ... More
EGR2 mutations in inherited neuropathies dominant-negatively inhibit myelin gene expression.
AuthorsNagarajan R, Svaren J, Le N, Araki T, Watson M, Milbrandt J
JournalNeuron
PubMed ID11394999
The identification of EGR2 mutations in patients with neuropathies and the phenotype Egr2/Krox20(-/-) have demonstrated that the Egr2 transcription factor is critical for peripheral nerve myelination. However, the mechanism by which these mutations cause disease remains unclear, as most patients present with disease in the heterozygous state, whereas Egr2(+/-) mice ... More
In vivo identification of ribonucleoprotein-RNA interactions.
AuthorsZielinski J, Kilk K, Peritz T, Kannanayakal T, Miyashiro KY, Eiríksdóttir E, Jochems J, Langel U, Eberwine J
JournalProc Natl Acad Sci U S A
PubMed ID16432185
To understand the role of RNA-binding proteins (RBPs) in the regulation of gene expression, methods are needed for the in vivo identification of RNA-protein interactions. We have developed the peptide nucleic acid (PNA)-assisted identification of RBP technology to enable the identification of proteins that complex with a target RNA in ... More
A role for a rat homolog of staufen in the transport of RNA to neuronal dendrites.
AuthorsTang SJ, Meulemans D, Vazquez L, Colaco N, Schuman E
JournalNeuron
PubMed ID11709157
RNAs are present in dendrites and may be used for local protein synthesis in response to synaptic activity. To begin to understand dendritic RNA targeting, we cloned a rat homolog of staufen, a Drosophila gene that participates in mRNA targeting during development. In hippocampal neurons, rat staufen protein displays a ... More
Multiplex cytological profiling assay to measure diverse cellular states.
AuthorsGustafsdottir SM, Ljosa V, Sokolnicki KL, Anthony Wilson J, Walpita D, Kemp MM, Petri Seiler K, Carrel HA, Golub TR, Schreiber SL, Clemons PA, Carpenter AE, Shamji AF
Journal
PubMed ID24312513
Computational methods for image-based profiling are under active development, but their success hinges on assays that can capture a wide range of phenotypes. We have developed a multiplex cytological profiling assay that "paints the cell" with as many fluorescent markers as possible without compromising our ability to extract rich, quantitative ... More
A double-label technique that monitors sulfur mustard damage to nuclei and mitochondria of normal human epidermal keratinocytes in vitro.
AuthorsCook JR, Van Buskirk RG
JournalToxicol Pathol
PubMed ID9323838
Sulfur mustard and 2-chloro ethyl ethyl sulfide (CEES, a sulfur mustard analog) is known to have immediate (minutes), long-term (hours to days), and toxic effects on human skin. Research was directed toward developing a single in vitro assay that might reflect both these short-term and long-term effects of this vesicating ... More
Colocalization of single ribosomes with intermediate filaments in puromycin-treated and serum-starved mouse embryo fibroblasts.
Previous experiments have revealed a relatively weak electrostatic binding capacity of in vitro reconstituted intermediate filaments (IFs) as well as of natural IFs of whole cell mount preparations for purified ribosomal particles of mammalian origin. In order to demonstrate that such associations also occur in vivo, intact cells were subjected ... More
Targeting G-quadruplex DNA as cognitive function therapy for ATR-X syndrome.
AuthorsShioda N, Yabuki Y, Yamaguchi K, Onozato M, Li Y, Kurosawa K, Tanabe H, Okamoto N, Era T, Sugiyama H, Wada T, Fukunaga K
JournalNat Med
PubMed ID29785027
'Alpha-thalassemia X-linked intellectual disability (ATR-X) syndrome is caused by mutations in ATRX, which encodes a chromatin-remodeling protein. Genome-wide analyses in mouse and human cells indicate that ATRX tends to bind to G-rich sequences with a high potential to form G-quadruplexes. Here, we report that Atrx mutation induces aberrant upregulation of ... More
Stress routes clients to the proteasome via a BAG2 ubiquitin-independent degradation condensate.
Authors
JournalNat Commun
PubMed ID35654899
Bioactivity screening of environmental chemicals using imaging-based high-throughput phenotypic profiling.
AuthorsNyffeler J, Willis C, Lougee R, Richard A, Paul-Friedman K, Harrill JA
JournalToxicol Appl Pharmacol
PubMed ID31899216
'The present study adapted an existing high content imaging-based high-throughput phenotypic profiling (HTPP) assay known as "Cell Painting" for bioactivity screening of environmental chemicals. This assay uses a combination of fluorescent probes to label a variety of organelles and measures a large number of phenotypic features at the single cell ... More
β1 integrin mediates an alternative survival pathway in breast cancer cells resistant to lapatinib.
Authors
JournalBreast Cancer Res
PubMed ID21884573
TFEB Transcriptional Responses Reveal Negative Feedback by BHLHE40 and BHLHE41.
Authors
JournalCell Rep
PubMed ID33176151
A phenomics approach for antiviral drug discovery.
Authors
JournalBMC Biol
PubMed ID34334126
The Galapagos Chip Platform for High-Throughput Screening of Cell Adhesive Chemical Micropatterns.
Authors
JournalSmall
PubMed ID34985808
Integrating deep learning and unbiased automated high-content screening to identify complex disease signatures in human fibroblasts.
Authors
JournalNat Commun
PubMed ID35338121
Natural product fragment combination to performance-diverse pseudo-natural products.
Authors
JournalNat Commun
PubMed ID33767198
Control of osteocyte dendrite formation by Sp7 and its target gene osteocrin.
Authors
JournalNat Commun
PubMed ID34725346
Cell Painting, a high-content image-based assay for morphological profiling using multiplexed fluorescent dyes.
AuthorsBray MA, Singh S, Han H, Davis CT, Borgeson B, Hartland C, Kost-Alimova M, Gustafsdottir SM, Gibson CC, Carpenter AE
JournalNat Protoc
PubMed ID27560178
In morphological profiling, quantitative data are extracted from microscopy images of cells to identify biologically relevant similarities and differences among samples based on these profiles. This protocol describes the design and execution of experiments using Cell Painting, which is a morphological profiling assay that multiplexes six fluorescent dyes, imaged in ... More
BDNF elevates the axonal levels of hnRNPs Q and R in cultured rat cortical neurons.
Local translation plays important roles in the maintenance and various functions of axons, and dysfunctions of local translation in axons are implicated in various neurological diseases. Heterogeneous nuclear ribonucleoproteins (hnRNPs) are RNA binding proteins with multiple functions in RNA metabolism. Here, we identified 20 hnRNPs in the axons of cultured ... More