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Genetic reporter for live tracing fluid flow forces during cell fate segregation in mouse blastocyst development

ABSTRACT: We used a robut low input cell and high throughput in-situ ChIP method to emphasize regulation mechanism among E3.5 klf2, E3.5 klf4 and E12.5 EC klf2.

ORGANISM(S): Mus musculus

PROVIDER: GSE197167 | GEO | 2023/07/31

REPOSITORIES: GEO

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Genetic reporter for live tracing fluid flow forces during cell fate segregation in mouse blastocyst development [scRNA-seq]

Project description:We performed single-cell RNA-seq in cultured embryos under shaking and static condition using Klf2-H2Bmcherry mice to screen an effective force reporter.

2023-07-31 | GSE197168 | GEO

Genetic reporter for live tracing fluid flow forces during cell fate segregation in mouse blastocyst development

Project description:This SuperSeries is composed of the SubSeries listed below.

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A dual-fluorescence reporter in the Eomes locus for live imaging and medium-term lineage tracing.

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RNA sequencing of single blastocyst-stage embryos (E3.5) from zygotes which were injected with sperm sRNAs from normal or depressive male mice.

Project description:We demonstrated that sperm sRNAs contribute to paternal transmission of depression-like symptoms.To further investigated the mechanism by which sperm sRNAs contribute to depression inheritance. Since the early embryonic period represents a window of plasticity important for adult phenotypes, we injected sperm sRNAs from mice of normal(F0-Ctl) or stress induced depressive(F0-Dep)into zygotes and assessed transcriptional changes when the embryos developed to the E3.5 blastocyst stage (sRNA-Dep-E3.5 vs sRNA-Ctl-E3.5).

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KLF2-induced actin shear fibers control both alignment to flow and JNK signaling in vascular endothelium

Project description:The shear stress-induced transcription factor KrÃ¼ppel-like factor 2 (KLF2) confers anti-inflammatory properties to endothelial cells through inhibition of activator protein 1, presumably by interfering with MAPK cascades. To gain insight into the regulation of these cascades by KLF2, we used antibody arrays in combination with time-course mRNA micro-array analysis. No gross changes in MAPKs were detected, rather phosphorylation of actin cytoskeleton-associated proteins, including Focal Adhesion Kinase, was markedly repressed by KLF2. Furthermore, we demonstrate that KLF2-mediated inhibition of Jun NH2-terminal kinase (JNK) and its downstream targets ATF2/c-Jun is dependent on the cytoskeleton. Specifically, KLF2 directs the formation of typical short basal actin filaments, we term shear fibers, which are distinct from thrombin- or TNF-Î±-induced stress fibers. KLF2 is shown to be essential for shear stress-induced cell alignment, concomitant shear fiber assembly and inhibition of JNK signaling. These findings link the specific effects of shear-induced KLF2 on endothelial morphology to the suppression of JNK MAPK signaling in vascular homeostasis via novel actin shear fibers. Tramscriptome profiling: Three independent isolates of Human Umbilical Vein Endothelial cells were transduced with lentiviral vectors expressing Kruppel Like Factor 2 (KLF2) or no protein (mock), and at time after transduction 24 h, 48 h, 72 h , RNA was isolated and hybridized to GPL4868 microarrays using dye swap procedure Kinome profiling: Two independent isolates of Human Umbilical Vein Endothelial cells were transduced with lentiviral vectors expressing Kruppel Like Factor 2 (KLF2) or no protein (mock), and at time after transduction 72 h , total cellular protein was isolated and hybridized to Kinexus KAM-1.1 phosphoprotein (kinexus) microarrays using dual color procedure in duplicate

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RNA sequencing of mouse embryonic day 3.5 blastocyst inner cell mass cells

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2014-05-15 | E-MTAB-2555 | biostudies-arrayexpress

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Bulk and single-cell RNA-seq of mouse blastocyst, blastoid Trophoblast stem cells and Embryonic stem cells

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