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KLF4 Time Course

ABSTRACT: A stably transfected inducible system for KLF4 was established in RKO cells as previously described (Chen et al., 2001; Chen et al., 2003). This cell line, called EcR-RKO/KLF4, contains stably transfected receptors for the insect hormone, ecdysone (EcR), and a full-length mouse KLF4 cDNA driven by a promoter that responds to ecdysone and its analogue, ponasterone A. Cells were treated with 5 µM ponasterone A (PA), for various time durations. To control for the experiment, the vehicle, ethanol, was added for the same periods of time. Keywords: Time course

ORGANISM(S): Homo sapiens

PROVIDER: GSE3113 | GEO | 2005/08/13

SECONDARY ACCESSION(S): PRJNA93121

REPOSITORIES: GEO

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Transcription profiling of human RKO cells after induction of KLF4 for different lengths of time

Project description:A stably transfected inducible system for KLF4 was established in RKO cells as previously described (Chen et al., 2001; Chen et al., 2003). This cell line, called EcR-RKO/KLF4, contains stably transfected receptors for the insect hormone, ecdysone (EcR), and a full-length mouse KLF4 cDNA driven by a promoter that responds to ecdysone and its analogue, ponasterone A. Cells were treated with 5 ÂµM ponasterone A (PA), for various time durations. To control for the experiment, the vehicle, ethanol, was added for the same periods of time.

2007-06-21 | E-GEOD-3113 | biostudies-arrayexpress

Opportunistic binding of EcR to open chromatin drives tissue-specific developmental responses [RNA-seq]

Project description:In Drosophila, the ecdysone steroid hormone is essential for coordinating developmental timing across physically separated tissues. Ecdysone directly impacts genome function through its receptor, a heterodimer of the EcR and Usp proteins. Ligand binding to EcR triggers a transcriptional cascade, including activation of a set of primary response transcription factors. The hierarchical organization of this pathway has left the direct role of EcR in mediating ecdysone responses obscured. Here, we investigate the role of EcR in controlling tissue-specific ecdysone responses, focusing on two tissues that diverge in their response to rising ecdysone titers: the larval salivary gland, which undergoes programmed destruction, and the wing imaginal disc, which initiates metamorphosis. We find that EcR functions bimodally, with both gene repressive and activating functions, even at the same developmental stage. We find that EcR DNA binding profiles are highly tissue-specific, and transgenic reporter analyses demonstrate that EcR plays a direct role in controlling enhancer activity. Despite a strong correlation between tissue-specific EcR binding and tissue-specific open chromatin, we find that EcR does not control chromatin accessibility at genomic targets. We conclude that EcR contributes extensively to tissue-specific ecdysone responses. However, control over access to its binding sites is subordinated to other transcription factors.

2022-09-12 | GSE202809 | GEO

Opportunistic binding of EcR to open chromatin drives tissue-specific developmental responses [FAIRE-seq]

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Opportunistic binding of EcR to open chromatin drives tissue-specific developmental responses [CUT&RUN]

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Direct and widespread role for the nuclear receptor EcR in mediating the response to ecdysone in Drosophila

Project description:Ecdysone signaling has long been studied as a paradigm for how hormones can synchronize development events in tissues throughout the animal. At the genetic level, ecdysone acts through its receptor, EcR, which has been classically thought to act at a relatively small number of canonical, ecdysone-responsive genes which then go on to activate the effectors of cellular processes. However, in spite of the decades of work studying EcR, the understanding of how EcR promotes changes in gene expression genome-wide in vivo remains incomplete. To examine EcR’s role in promoting changes in gene expression, we use a tissue-specific GAL4 driver to knockdown EcR throughout wing development and perform RNAseq on both WT wings and EcR-RNAi wings at -6h after puparium formation (APF) and +6hAPF. We find that EcR is required to promote thousands of gene expression changes genome-wide. To determine whether EcR is directly effecting these changes, we perform CUT&RUN in wings to identify EcR binding sites genome-wide. We find that EcR directly binds thousands of sites genome-wide, including many genes important for wing development. Collectively, these data suggest that EcR does not only act the top of a signaling cascade, but, instead, directly effects the response to ecdysone at multiple levels.

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Epigenome analysis of control and KLF4-overexpressing podocytes

Project description:Genome wide DNA methylation profiling of control (vector-transfected) and KLF4-overexpressing human podocytes (parental cell line: Saleem et al. JASN 13:630, 2002). The Illumina HumanMethylation450 Beadchip was used to obtain DNA methylation profiles.

2014-11-01 | GSE41689 | GEO

Drosophila Ecdysone Receptor Study

Project description:We used the DamID method to systematically identify the binding sites of Ecdysone Receptor and its heterodimeric partner USP across the whole genome in Drosophila Kc cells. We find that the EcR sites are a subset of the USP sites and that only a proportion are ecdysone regulated from an accompanying ecdysone profiling study. The role of EcR/USP in the ecdysone network appears to be coordinated by the recruitment of many transcription factors as well as signaling molecules. Keywords: DamID, chromatin profiling, DNA microarray

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EcR mutant midgut comparison

Project description:Comparisons between EcR mutant midgut and a reference control sample from mixed-stage normal midgut Keywords = Drosophila, ecdysone, network, genomic, microarray, organogenesis, EcR, midgut, central nervous system, salivary gland, epidermis, imaginal disc, development

2010-06-05 | E-GEOD-540 | biostudies-arrayexpress

EcR mutant midgut comparison

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Cytokinesis signals truncation of the PodJ polarity factor by a cell cycle-regulated protease

Project description:Supporting data for Chen et al., "Cytokinesis signals truncation of the PodJ polarity factor by a cell cycle-regulated protease" The microarray component of this work seeks to identify genes whose expression is controlled by the Caulobacter crescentus DivJ-PleC-DivK system. Keywords: time course, mutant/wild-type comparisons

2005-12-08 | GSE3144 | GEO

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