Project description:A Flag-tagged version of murine EPOP was expressed in mouse embryonic stem cells (ESCs) at near to endogenous levels and purified using an anti-FLAG affinity matrix.
Project description:A Flag-tagged version of murine Elongin B was expressed in mouse embryonic stem cells (ESCs) at near to endogenous levels and purified using an anti-FLAG affinity matrix.
Project description:We found that the non-essential amino acid L-Proline (L-Pro) acts as a signaling molecule that promotes the conversion of embryonic stem cells (ESCs) into mesenchymal-like, spindle-shaped, highly motile, invasive pluripotent stem cells. This embryonic stem cell-to-mesenchymal-like transition (esMT) is accompanied by a genome-wide remodeling of the transcriptome We used microarrays to elucidate whether a diverse transcriptional program is the basis of the morphological and motility differences between ESCs and L- Proline treated ESCs (PiCs) Total RNA was extracted from Control (ESCs) and L-Proline treated mouse embryonic stem cells (PiCs) and hybridized on Affimetrix microarrays.
Project description:Sin3a, a known master scaffold, provides unique contact surfaces for interaction with particular accessory proteins to repress the transcription of specific genes. Surprisingly, our results also suggest that Sin3a has a role in transcriptional activation. We compared gene expression differences between Sin3a knockdown and control ESCs with mouse gene expression microarrays and identified an approximately equal distribution of up- and down-regulated genes following Sin3a knockdown in mouse ESCs. We propose that Sin3a collaborates with Tet1 to demethylate adjacent genomic regions and, ultimately, facilitates actively transcribed gene expression in mouse ESCs.
Project description:TET1 maintains hypomethylation at bivalent promoters through its catalytic activity in embryonic stem cells (ESCs). However, whether and how TET1 exerts catalytic activity-independent functions in regulating bivalent genes is not well understood. Therefore, we mapped the TET1 interactome in mouse ESCs using a SILAC IP-MS proteomics approach.
Project description:In pluripotent mouse embryonic stem cells (ESCs), SetDB1, which targets up to twenty thousand genomic loci, plays an essential role in regulating various cellular processes, including maintenance of the embryonic stem cell state, differentiation and metabolism. SetDB1 is best known as a chromatin modifier catalyzing H3K9me3. However, recent studies show that SetDB1 can mediate H3K27me3-deposition by PRC2, CTCF-binding, recruit DNA methyltransferases, and potentially regulate additional chromatin processes. To gain further insights, we profiled the chromatin-bound protein landscapes of WT mouse ESCs, as well as SetDB1-deficient mouse ESCs at different time points after inducing Setdb1 KO, using Chromatin Enrichment for Proteomics (ChEP + LC-MS/MS).
Project description:We found that the non-essential amino acid L-Proline (L-Pro) acts as a signaling molecule that promotes the conversion of embryonic stem cells (ESCs) into mesenchymal-like, spindle-shaped, highly motile, invasive pluripotent stem cells. This embryonic stem cell-to-mesenchymal-like transition (esMT) is accompanied by a genome-wide remodeling of the transcriptome We used microarrays to elucidate whether a diverse transcriptional program is the basis of the morphological and motility differences between ESCs and L- Proline treated ESCs (PiCs)
Project description:We found that the non-essential amino acid L-proline (L-Pro) acts as a signaling molecule that promotes the conversion of embryonic stem cells (ESCs) into mesenchymal-like, spindle-shaped, highly motile, invasive pluripotent stem cells.This embryonic stem cell-to-mesenchymal-like transition (esMT) is accompanied by a genome-wide remodeling of the H3K9me3 and H3K36me3 histone marks. Examination of 2 different histone modifications in untreated ESCs and L-Pro treated ESCs