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1-deoxysphingolipids bind to COUP-TF to modulate lymphatic and cardiac cell development

ABSTRACT: We identified 1-deoxysphingosine as a ligand for NR2F1 and NR2F2. To prove physiological relevance after in vitro binding assay we investigated the role of NR2F1 and NR2F2 in cardiomyocyte differentiation. We also used different strategies to manipulate levels of 1-deoxysphingosines and canonical sphingosines in cardiomyocytes derived from human embryonic stem cells. Here we generated both single knockout of NR2F2 and double knockouts of NR2F1 and NR2F2 in human embryonic stem cell line H9. The differentiation of WT, KO and DKO lines into cardiomyocytes was then compared by single-cell RNA-seq analysis.

ORGANISM(S): Homo sapiens

PROVIDER: GSE185456 | GEO | 2021/10/08

REPOSITORIES: GEO

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1-deoxysphingolipids bind to COUP-TF to modulate lymphatic and cardiac cell development

Project description:We identified 1-deoxysphingosine as a ligand for NR2F1 and NR2F2. To prove physiological relevance after in vitro binding assay we used different strategies to manipulate levels of 1-deoxysphingosines and canonical sphingosines in cardiomyocytes derived from human embrynonic stem cells. These manipulations included treating cells with sphingolipid biosynthesis inhibitors (myriocin), adding 1-deoxysphingosines to culture media or induced expression of the Sptlc1 HSAN mutant C133W to enhance 1-deoxysphingosine synthesis by cardiomyocytes.

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Atrial Identity Is Determined by A COUP-TFII Regulatory Network (ChIP-Seq)

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Atrial Identity Is Determined by A COUP-TFII Regulatory Network (ChIP-Seq)

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Genome-wide analysis of p300, H3K4me3, H3K4me1, H3K27me3 and H3K27ac in in vitro differentiated human neural crest cells (hNCC)

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Anti-NR2F1 scCUT&Tag on FAC-sorted Isl1MN-GFP rhombomere 4 neurons from wildtype and HCFP1 Fam5snv/snv E10.5 mouse embryos

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Schwann cell regulation by NR2F2 using Cut&Run

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RNAi profiling of human lymphatic endothelial cells after knockdown of Prox1 and/or NR2F2

Project description:Gene expression profiles of primary lymphatic endothelial cells (LECs) isolated from human foreskin were analyzed after siRNA-mediated knockdown of control (firefly luciferase), Prox1, NR2F2 or Prox1/NR2F2 for 48 hours. Experiment Overall Design: Passage five human lymphatic endothelial cells (LECs) were cultured on fibronectin (10 Î¼g/ml)-coated plates in a complete media (EBM, 20% FBS supplemented with 10 Î¼g/ml hydrocortisone acetate, 25 ug/ml cAMP and antibiotics). LECs were harvested and electorporated with siRNA duplexes for 48 hours with siRNA duplexes against either firefly luciferase(control), Prox1, NR2F2 or Prox1/NR2F2. Total RNA was purified using Tri-reagent and was subjected to microarray analysis. Experiment Overall Design:

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RNA-seq experiments in human neural crest cells (hNCC)

Project description:Combining an in vitro hNCC differentiation protocol with epigenomic profiling, we provide the first whole-genome characterization of cis-regulatory elements in this highly relevant cell type. With this data at hand, we have characterized the chromatin state and dynamics of all human gene promoters during the course of NCC in vitro differentiation. Most importantly, we have identified a large cohort of active and NCC-specific enhancers, which we showed to be functionally relevant in vivo, in the context of embryonic development. Finally, through sequence analysis of the identified NCC enhancers, we uncovered the orphan nuclear receptors NR2F1 and NR2F2 as novel hNCC transcriptional regulators both in vitro and in vivo. RNA-seq experiments in human neural crest cells (hNCC)

2012-09-13 | E-GEOD-28875 | biostudies-arrayexpress

Epigenomic profiling of enhancers identifies orphan nuclear receptors NR2F1 and NR2F2 as novel regulators of human neural crest.

Project description:Combining an in vitro hNCC differentiation protocol with epigenomic profiling, we provide the first whole-genome characterization of cis-regulatory elements in this highly relevant cell type. With this data at hand, we have characterized the chromatin state and dynamics of all human gene promoters during the course of NCC in vitro differentiation. Most importantly, we have identified a large cohort of active and NCC-specific enhancers, which we showed to be functionally relevant in vivo, in the context of embryonic development. Finally, through sequence analysis of the identified NCC enhancers, we uncovered the orphan nuclear receptors NR2F1 and NR2F2 as novel hNCC transcriptional regulators both in vitro and in vivo. Genome-wide analysis of H3K27ac in chicken neural crest cells (NCC) obtained from stage 11-14 or stage 20 embryos

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