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HESC MIXL1+ MIXL1- microarray

ABSTRACT: Microarray analysis of isolated hES cells from day 3 of cardiac differentiation was used to identify differences between MIXL1eGFP+ and MIXL1eGFP- transcriptomes. We identified 6,757 differentially regulated genes, of which 2,520 were upregulated ≥2-fold in the eGFP+ (MIXL1+) mesoderm population

ORGANISM(S): Homo sapiens

PROVIDER: GSE74664 | GEO | 2015/11/05

SECONDARY ACCESSION(S): PRJNA301126

REPOSITORIES: GEO

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hESC MIXL1+ MIXL1- microarray

Project description:Microarray analysis of isolated hES cells from day 3 of cardiac differentiation was used to identify differences between MIXL1eGFP+ and MIXL1eGFP- transcriptomes. We identified 6,757 differentially regulated genes, of which 2,520 were upregulated â¥2-fold in the eGFP+ (MIXL1+) mesoderm population A stencil differentiation protocol was used to isolate mesodermal cells based on GFP expression from the MIXL1 locus. Microarray analysis of isolated cells from day 3 of differentiation was used to identify differences between MIXL1eGFP+ and MIXL1eGFP- transcriptomes.

2015-11-05 | E-GEOD-74664 | biostudies-arrayexpress

Change in Expression in U937 under MIXL1 enforced overexpression

Project description:While the role of the paired-type homeobox transcription factor MIXL1 in early embryonic development has been previously established, very little is known about the role of MIXL1 in adult hematopoiesis. To determine what role MIXL1 plays in adult hematopoietic cells, we established an enforced expression model for MIXL1 in the cell line U937, then analyzed the changes of gene expression caused by increased expression of MIXL1 by microarray analysis.

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Identification of targetted gene loci by MIXL1 in the AML cell line U937

Project description:While the role of the paired-type homeobox transcription factor MIXL1 in early ebryonic development has been previously established, very little is known about the role of MIXL1 in adult hematopoiesis. To determine what role MIXL1 plays in adult hematopoietic cells, we used an established enforced expression line for FLAG-tagged MIXL1 in U937, then isolated targetted regions of the genome by ChIP-Seq. Through this analysis, we identified new putative direct transcirptional targets for MIXL1 in AML and hematopoiesis.

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Single-cell RNAseq of Mixl1 knockout chimeric mouse embryos at embryonic day 8.5 of mouse development.

Project description:Chimeric embryos were generated to investigate the effect of Mixl1 knockout in mouse embryos by single-cell RNA-sequencing. Mixl1 is an essential transcription factor for mesendoderm development. Chimeric embryos contain tissue that is Mixl1+/+, which prevents global developmental failures. Embryos were generated by blastocyst injection of tdTomato-labelled, Mixl1-/- mouse embryonic stem cells into wild type embryos. After blastocyst harvest, cells were flow-sorted before 10X Genomics library preparation and single-cell RNA-sequencing.

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Gene expression analysis of hESC-derived-MESP1 expressing progenitors

Project description:A hESC MESP1-MCHERRY reporter line was used to isolate and study the molecular character of MESP1 expressing pre-cardiac progenitors, derived from hESC. MESP1 is a key-transcription factor for pre-cardiac mesoderm and is marking the progenitor for almost all cells of the heart. This reporter line was used to study cardiac differentiation and the derivation of early cardiac progenitors in vitro. hESCs were differentiated towards the cardiac lineage, expressing MESP1-mCherry at day 3 of differentiation. Total RNA obtained from isolated MESP1-mCherry expressing progenitors was compared to that of non-MESP1-expressing progenitors and undifferentiated hESCs in order to characterize MESP1-specific transcription factors and proteins.

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TMEM88 knockdown during human embryonic stem cell cardiac differentiation

Project description:TMEM88 is indispensable for heart development and acts in the pre-cardiac mesoderm to specify lineage commitment of the cardiovascular progenitor cell through inhibition of Wnt signaling. 2 different lentiviruses expression shRNA targeting different domains of the TMEM88 locus were transduced into undifferentiated hES cells. Cells were puromycin selected then differentiated along the cardiac lineage. Total RNA was taken at day 5 of differentiation when the cardiovascular progenitor cell arises. 1 sample for each shRNA and 2 samples for control shRNA were analyzed by array

2013-10-21 | E-GEOD-43805 | biostudies-arrayexpress

YAP Repression of the WNT3 Gene Controls hESC Differentiation Along the Cardiac Mesoderm Lineage

Project description:In hESCs, Wnt3/β-catenin activity is low and Activin/SMAD signaling ensures NANOG expression to sustain pluripotency. In response to exogenous Wnt3 effectors, Activin/SMADs switch to cooperate with β-catenin and induce mesendodermal differentiation genes. We show here that the HIPPO effector YAP binds to the WNT3 gene enhancer and prevents the gene from being induced by Activin in proliferating hESCs. In the absence of YAP, Activin signaling is sufficient to induce expression of the endogenous Wnt3 cytokine, which stabilizes β-catenin and selectively activates genes required for cardiac mesoderm (ME) formation. Interestingly, Activin-stimulated YAP-knockout hESCs strongly express β-catenin-dependent cardiac mesoderm markers (BAF60c and HAND1), but unlike WT hESCs, fail to express cardiac inhibitor genes (CDX2, MSX1). Accordingly, YAP-/- cells treated with Activin alone can differentiate efficiently to beating cardiomyocytes in culture, bypassing the need for sequential treatment with exogenous Wnt ligand and Wnt inhibitors. Similarly, Activin in combination with small-molecule YAP inhibitors generates beating cardiomyocytes from wild-type hESCs following a one- step protocol. Our findings highlight an unanticipated role of YAP as an upstream regulator of WNT3 to maintain hESC pluripotency in the presence of Activin, and uncover a direct route for the development of human embryonic cardiac mesoderm.

2018-01-08 | GSE99202 | GEO

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