ABSTRACT: Here we performed a RNA-seq experiment on samples of adherent cultures of mouse neural stem cells (NS5 cell line) under normal growth conditions and after 4 hours of treatment with the gamma-secretase inhibitor LY 411575. This resulted in the generation of a genome-wide mRNA expression pattern and quantification for these cells in the two conditions.
Project description:Here we performed a microarray experiment on samples of adherent cultures of mouse neural stem cells (NS5 cell line) expressing an inducible version of the transcription factor MyT1 (MyT1-V5 TetON) under normal growth conditions and after 4 hours of treatment by doxycyline. This resulted in the generation of a genome-wide mRNA expression pattern and quantification for these cells in the two conditions.
Project description:Here we performed a ChIP-seq experiment on a sample of adherent cultures of mouse neural stem cells (NS5 cell line) expressing an inducible HA-tagged version of the proneural factor MyT1 (MyT1-HA, under TetON control) after activation by doxycycline hyclate (DOX). This resulted in the generation of a genome-wide map of MyT1-HA binding to chromatin.
Project description:Here we performed a ChIP-seq experiment on a sample of adherent cultures of mouse neural stem cells (NS5 cell line) under normal growth conditions and upon short term activation (30 minutes) of an inducible version of the proneural factor Mash1/Ascl1 (Ascl1-ERT2). This resulted in the generation of a genome-wide map of Ascl1 binding to chromatin.
Project description:Here we performed a ChIP-seq experiment on a sample of adherent cultures of mouse neural stem cells (NS5 cell line) expressing an inducible version of the proneural factor Mash1/Ascl1 (Ascl1-ERT2) under normal growth conditions and after 24 hours of activation by 4-Hydroxytamoxifen. This resulted in the generation of a genome-wide map of histone modifications H3K27ac and H3K4me1.
Project description:Here we performed a ChIP-seq experiment on a sample of adherent cultures of mouse neural stem cells (NS5 cell line) under normal growth conditions. This resulted in the generation of a genome-wide map of Zeb1 binding to chromatin.
Project description:Here we performed a ChIP-seq experiment on a sample of adherent cultures of mouse neural stem cells (NS5 cell line) under normal growth conditions. This resulted in the generation of a genome-wide map of RBPJ binding to chromatin.
Project description:The incorporation of histone H3 variants has been implicated in the epigenetic memory of cellular state. Using genome editing with zinc finger nucleases to tag endogenous H3.3, we report genome-wide profiles of H3 variants in mammalian embryonic stem (ES) cells and neuronal precursor cells. Genome-wide patterns of H3.3 are dependent on amino acid sequence, and change with cellular differentiation at developmentally regulated loci. The H3.3 chaperone Hira is required for H3.3 enrichment at active and repressed genes. Strikingly, Hira is not essential for localization of H3.3 at telomeres and many transcription factor binding sites. Immunoaffinity purification and mass spectrometry reveal that the proteins Atrx and Daxx associate with H3.3 in a Hira-independent manner. Atrx is required for Hira-independent localization of H3.3 at telomeres, and for the repression of telomeric RNA. Our data demonstrate that multiple and distinct factors are responsible for H3.3 localization at specific genomic locations in mammalian cells. Crosslinking ChIP-seq: Examination of 1 histone variant (H3.3), 2 histone modifications, and Serine-5 phosphorylated RNA polymerase in 2 different cell types (H3.3-HA ES samples 1-4, and H3.3-HA NPC samples 7-10). Examination of 1 histone variant (H3.2), and one histone modification (H3K36me3) in 2 different cell types (H3.2-HA ES samples 5-6, and H3.2-HA NPC samples 11-12). Examination of 1 histone variant (H3.3), input control, and one histone modification (H3K36me3) in one cell type (H3.3-HA hybrid ES, samples 13-15). Examination of 1 histone variant (H3.1S31), input control, and one histone modification (H3K36me3) in one cell type (H3.1S31-HA hybrid ES, samples 16-18). Native ChIP-seq: Examination of 1 histone variant (H3.3), input control, and one histone modification (H3K4me3) in one cell type (H3.3-HA ES, samples 19-21). Examination of 1 histone variant (H3.2), input control, and two histone modifications (H3K4me3 and H3K27me3) in one cell type (H3.2-HA ES, samples 22-25). Examination of 1 histone variant (H3.3), input control, and two histone modifications (H3K4me1 and H3K36me3) in one cell type (H3.3-EYFP ES, samples 26-29). Examination of 1 histone variant (H3.3), input control, and two histone modifications (H3K4me1 and H3K36me3) in one cell type (Hira -/- H3.3-EYFP ES, samples 30-33). Examination of 1 histone variant (H3.3) and input control in one cell type (Atrxflox H3.3-EYFP ES, samples 34-37). Examination of HA antibody background in one cell type (wild-type ES, sample 38).
Project description:The ability of high-risk neuroblastoma to survive unfavorable growth conditions and multimodal therapy is hypothesized to result from a phenomenon known as reversible adaptive plasticity (RAP). RAP is a novel phenomenon enabling neuroblastoma cells to transition between a proliferative anchorage dependent (AD) state and a slow growing anoikis-resistant anchorage independent (AI) state. We used microarrays to investigate the global gene expression profiles in AD and AI cells, and to identify the differential expressed genes within signaling pathways contributing to the reversible adaptive plasticity between AD and AI cells. Comparison of microarray data from AD cells (n=4 independent cultures) versus AI cells (n=4 independent cultures) were performed using Partek Genomics Suite 6.5. Differentially expressed genes with an FDR M-bM-^IM-$5% and a fold-change M-bM-^IM-%1.5 were selected for pathway analysis.
Project description:Among the flaviviral proteins, NS5 is the largest and most conserved. NS5 contains major enzymatic components of the viral replication complex. Disruption of the common key NS5-host protein-protein interactions critical for viral replication could aid in the development of broad-spectrum anti-flaviviral therapeutics. To this end, we investigated the JEV- and ZIKV-NS5 interactomes in human cells using GFP pull-downs with mass spectrometry analysis in a label-free fashion. A total of 138 cellular proteins interacting with NS5 from JEV, ZIKV, or both were identified as Protein classification analysis of identified cellular targets revealed the enrichment of RNA binding, processing and splicing including spliceosomal and spliceosome-associated proteins in both datasets. Comparison of our data with literature not only revealed several cellular NS5 interacting proteins shared among flaviviruses, but also identified proteins that have no known function in flavivirus biology such as RNA polymerase II-associated Paf1 complex, protein phosphatase 6, and s-adenosylmethionine synthetase. Our study generates the first landscape of the JEV and ZIKV NS5 interactome in human cells and identifies cellular proteins that are potentially targetable for broad-spectrum anti-flaviviral therapy.