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A Myc-dependent role for cohesin in the heterokaryon-mediated reprogramming of somatic nuclei

ABSTRACT: Analysis of gene expression changes in mESCs where the cohesin subunit Scc1 has been depleted. There are 6 samples. 3 Scc1 wt/wt ERT2-Cre biological replicas and 3 Scc1 lox/lox ERT2-Cre biological replicas, all treated with 4'-OHT 80nM final concentration for 24h.

ORGANISM(S): Mus musculus

PROVIDER: GSE62325 | GEO | 2015/02/19

SECONDARY ACCESSION(S): PRJNA263831

REPOSITORIES: GEO

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A Myc-dependent role for cohesin in the heterokaryon-mediated reprogramming of somatic nuclei

Project description:Analysis of gene expression changes in mESCs where the cohesin subunit Scc1 has been depleted. There are 6 samples. 3 Scc1 wt/wt ERT2-Cre biological replicas and 3 Scc1 lox/lox ERT2-Cre biological replicas, all treated with 4'-OHT 80nM final concentration for 24h. Scc1 wt/wt and Scc1 lox/lox ERT2-Cre mES cells where grown in N2B27 +2i+LIF medium (Ying et al. 2008) for 24h in the presence of 80mM 4-OHT to induce cohesin depletion. Three biological replicates were collected and RNA was extracted. Microarray samples were prepared following the manufacturers instructions (Affymetrix) using 200ng of total RNA. Microarry samples were hybridized to Affymetrix GeneChip Mouse Gene 1.0 ST Arrays and the analysis was performed using the Bioconductor Package for R.

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2016-05-18 | E-GEOD-72024 | biostudies-arrayexpress

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2022-03-10 | PXD030748 | Pride

Next Generation Sequencing (RNA-Sequencing) for the analysis of RUNX3 targets in H460, H460-ERT2-RUNX3 WT and H460-ERT2-RUNX3 MT(K94/171R mutation)

Project description:Purpose: The goal of this study is to compare the gene expression profiles of H460, H460-ERT2-RUNX3-WT and H460-ERT2-RUNX3-MT(K94/171R) cells. Methods: H460, H460-ERT2-RUNX3 WT, and H460-ERT2-RUNX3-MT(K94/171R) cells were serum-starved for 24 hr, and then stimulated with 10% serum or 10% serum + 1 uM 4-OHT for 0, 8, or 16 hr. RNA was extracted from the cells. Isolated total RNA was processed for preparation of an RNA-seq library using the Illumina TruSeq Stranded mRNA Sample Preparation kit (Illumina, San Diego, CA, USA). Quality and size of libraries were assessed using the Agilent 2100 Bioanalyzer DNA kit (Agilent, Santa Clara, CA, USA). All libraries were quantified by qPCR using a CFX96 Real Time System (Bio-Rad, Hercules, CA, USA) and sequenced on NextSeq500 sequencers (Illumina). Sequencing adapters and low-quality bases in the raw reads were trimmed using the Cutadapt software. The cleaned high-quality reads were mapped to the human reference genome hg19 (https://genome.ucsc.edu) using STAR software. Genes differentially expressed between two selected biological conditions were identified by Cuffdiff in the Cufflinks package (http://cole-trapnell-lab.github.io/cufflinks/papers/). Results: RNA-Sequencing Data suggest that the R-point defends against oncogenic K-RAS–induced tumorigenesis not only by regulating intracellular programs (cell cycle, apoptosis, and metabolic pathways), but also by regulating extracellular programs (inflammatory response and immune response).

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