The Ets family transcription factor PU.1 is essential for the development and maintenance of several hematopoietic lineages. In the thymus, PU.1 is expressed only in the early ETP/DN1, DN2a and DN2b stages of development. While PU.1 deletion in multipotent precursors leads to a complete block in T-cell development its function in the intrathymic stages in which it is expressed remains undetermined. The goal of this expression profiling study was to determine if PU.1 regulates the expression of T-lineage genes during the early stages of development. To do this, we generated the PU.1-Eng construct which expresses a fusion protein containing the DNA binding ETS domain of PU.1 (aas 159-260) fused to the obligate repressor domain (aas 1-298) of the Drosophila engrailed protein. The PU.1-ETS construct only expresses the ETS domain of PU.1 (aas 159-260) and serves as a control. Fetal liver precursors were isolated from e14.5 embryos and co-cultured with OP9-DL1 cells in the presence of IL-7 and Flt3L (5 ng/ml each) for 4 days to obtain FLDN1, DN2a and DN2b cells. These were infected with vector only, PU.1-ETS and the PU.1-Eng constructs and DN2 cells were sorted after 20 hours of infection. Total RNA was isolated from these cells and polyA+ fraction was used to prepare libraries for high throughput sequencing. Libraries prepared from 2 independent sets of samples were subjected to non-strand specific single-end sequencing. Overall design: Two sets of samples generated from fetal liver precursor derived DN2 cells expressing PU.1-ETS and PU.1-Eng constructs were used for expression profiling. The LZRS retroviral vector, without any insert, was used to generate the vector control dataset.
Regulation of early T-lineage gene expression and developmental progression by the progenitor cell transcription factor PU.1.
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HNF6 and Rev-erbα integrate hepatic lipid metabolism by overlapping and distinct transcriptional mechanisms.
Sex, Age, Specimen part
View SamplesWe address the function of HNF6 in the mouse liver metabolism and Rev-erba cistrome
HNF6 and Rev-erbα integrate hepatic lipid metabolism by overlapping and distinct transcriptional mechanisms.
Sex, Age, Specimen part
View SamplesDuring T cell development, multipotent progenitors relinquish competence for other fates and commit to the T cell lineage by turning on Bcl11b, which encodes a transcription factor. To clarify lineage commitment mechanisms, we followed developing T cells at the single-cell level using Bcl11b knock-in fluorescent reporter mice. Notch signaling and Notch activated transcription factors collaborate to activate Bcl11b expression irrespectively of Notch-dependent proliferation. These inputs work via three distinct, asynchronous mechanisms: an early locus ‘poising’ function dependent on TCF-1 and GATA-3, a stochastic-permissivity function dependent on Notch signaling, and a separate amplitude-control function dependent on Runx1, a factor already present in multipotent progenitors. Despite their necessity for Bcl11b activation, these inputs act in a stage specific manner, providing a multitiered mechanism for developmental gene regulation. Overall design: Two sets of samples were generated from DN T-cell sub-populations derived from culture of bone marrow progenitors from mice containing a knock-in Bcl11b-YFP reporter
Asynchronous combinatorial action of four regulatory factors activates Bcl11b for T cell commitment.
Specimen part, Subject
View SamplesThe purpose of the study was to determine what genes in DN2 pro-T cells are immediately regulated by the transcription factor GATA-3, either as activation targets or as repression targets. To do this, two pairs of Gata3-floxed and control pro-T cells were generated and analyzed by RNA-seq within the first day of deletion of the Gata3 gene. Pro-T cells were generated by differentiation in vitro on OP9-DL1 monolayers of fetal liver-derive precursors from wildtype or Gata3-floxed mice, and the Gata3 gene was acutely deleted by transduction with Cre retroviral vector. Within 20 hr after transduction, samples of acutely Gata3-deleted and control DN2 cells were sorted and RNA prepared for RNA-seq analysis. High-throughput sequencing of the samples was carried out. Experimental Gata3 deleted samples in both cases were Gata3-floxed, ROSA26R-EYFP samples infected with Cre retrovirus and sorted for EYFP+ (Cre-activated) DN2 phenotype. Control for experiment 1: wildtype (C57BL/6) DN2 pro-T cells generated in parallel, also treated with Cre retrovirus but sorted only for DN2 phenotype. Control for experiment 2: same genotype as experimental, but infected with a GFP+ empty retroviral vector and sorted for GFP+ DN2 phenotype. Overall design: Two pairs of RNA-seq samples of DN2 pro-T cells were generated for comparison, each pair consisting of a Gata3-deleted sample plus a stage-matched control.
GATA-3 dose-dependent checkpoints in early T cell commitment.
No sample metadata fields
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The hepatic circadian clock fine-tunes the lipogenic response to feeding through RORα/γ.
Sex, Age, Specimen part
View SamplesWe address the function of ROR in the mouse liver metabolism
The hepatic circadian clock fine-tunes the lipogenic response to feeding through RORα/γ.
Sex, Age, Specimen part
View SamplesThis SuperSeries is composed of the SubSeries listed below.
GENE REGULATION. Discrete functions of nuclear receptor Rev-erbα couple metabolism to the clock.
Specimen part, Time
View SamplesGene expression analysis showed that LncPHx2 depletion resulted in upregulation of mRNAs encoding proteins known to promote cell proliferation, including MCM components, DNA polymerases, histone proteins, and transcription factors Overall design: RNA-seq analysis was performed on livers of mice subjected to PHx or to sham surgery after treatment with LncPHx2_ASO1 or with PBS. Gene expression profiling was done at 48 hours post-surgery
Partial Hepatectomy Induced Long Noncoding RNA Inhibits Hepatocyte Proliferation during Liver Regeneration.
No sample metadata fields
View SamplesCircadian and metabolic physiology are intricately intertwined, as illustrated by Rev-erb , a transcription factor (TF) that functions both as a core repressive component of the cell autonomous clock and as a regulator of metabolic genes. Here we show that Rev-erb modulates the clock and metabolism by different genomic mechanisms. Clock control requires Rev-erb to bind directly to the genome at its cognate sites, where it competes with activating ROR TFs. By contrast, Rev-erb regulates metabolic genes primarily by recruiting the HDAC3 corepressor to sites to which it is tethered by cell type-specific transcription factors. Thus, direct competition between Rev-erb and ROR TFs provides a universal mechanism for self-sustained control of molecular clock across all tissues, whereas Rev-erb utilizes lineage-determining factors to convey a tissue-specific epigenomic rhythm that regulates metabolism tailored to the specific need of that tissue.
GENE REGULATION. Discrete functions of nuclear receptor Rev-erbα couple metabolism to the clock.
Specimen part, Time
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