Project description:Study was carried out to investigates the acute responses of mouse ES cells to BMP-2. E14 cells were treated with BMP-2 at 25ng/ml for 1hour or untreated for the same time. The RNA were extracted and labeled to applied for microaaray.
Project description:In order to determine the impact of L3mbtl2 on genome wide mRNA expression microarray analysis was carried out in L3mbtl2 knock out ES cells and wild type ES cells. The same analysis was also carried out in knock out cells infected with L3mbtl2-F vector and a control vector.
Project description:Our previous studies have shown that the regulatory network that maintains pluripotency in mouse embryonic stem cells (mESCs) is regulated in a context-dependent manner and can be modulated, at least in part, by re-calibration of an intracellular network of pluripotency factors as well as cues arising from the extracellular matrix. The transcriptional repressor REST represses miR-21 and, thus, regulates self-renewal in E14Tg2a.4 mESCs cultured in the absence of mouse embryonic fibroblast feeder cell effects. However, how miR-21 connects to the nuclear regulatory network has not been clear. Here, we show that miR-21, a direct target of REST-mediated repression, directly targets Sox2. Exogenously added miR-21 to mESCs decreases the expression of Sox2, decreasing mESC self-renewal, and this effect of miR-21 on mESC self-renewal can be blocked by expression of exogenous Sox2. Conversely, destabilization of Sox2 by miR-21 can be blocked by anti-miR-21. Thus, the REST-miR-21-Sox2 axis connects REST to the core nuclear pluripotency regulators in E14Tg2a.4 mESCs cultured in the absence of feeder cells.
Project description:Carrying out both RNA-seq and Smad1/5 genome-wide chromatin immunoprecipitation and sequencing (ChIP-seq) analyses of mESCs in the naïve or primed states, we revisit the roles of BMP signaling in mESCs. RNA-seq analysis in 2 cell types; mESCs and ES-derived EpiSC (ESD-EpiSCs).
Project description:Carrying out both RNA-seq and Smad1/5 genome-wide chromatin immunoprecipitation and sequencing (ChIP-seq) analyses of mESCs in the naïve or primed states, we revisit the roles of BMP signaling in mESCs. Smad1/5 binding sites in 2 cell types; mESCs and ES-derived EpiSC (ESD-EpiSCs).
Project description:To determine how SAHA-PIP G can trigger the expression of the mesendoderm-related genes in mouse ES cells, chromatin immunoprecipitation (ChIP-seq) analysis was carried out with an antibody against the histone H3 lysine 4 trimethylation (H3K4me3) using EBs harvested on day 0
Project description:To identify proteins associated with mouse Elongin A (EloA), we carried out Co-IP on nuclear extract prepared from mouse ES cells from wild-type and null Elongin A cells (negative control).Each immunoprecipitation was excised as 5 gel
sections, minced and digested in-gel using trypsin. Electroionization was performed on the peptides before entering an LTQ Orbitrap Velos Pro ion trap mass spectrometer instrument (Thermo Fisher Scientific). Sequest (Thermo Finnigan, San Jose, CA) was used for identification of the peptide sequences. Identified peptides were filtered by false discovery rate of 1% or less. Processing, preparation, and Mass spectrometry analysis of the samples was carried out at Taplin Mass Spectrometry Facility (Harvard Medical School).
Project description:Recently, it was shown that the Bmp antagonist Noggin could strongly induce cardiomyocyte differentiation by transient treatment of undifferentiated ES cells. In order to determine how Noggin may induce cardiac differentiation, we compared differentially expressed genes during Noggin treatment of ES cells using microarray analysis and found that matrix metalloproteinase (Mmp)-3 is the only gene whose expression is increased by Noggin treatment. Keywords: embryonic stem cells, cardiac differentiation, matrix metalloproteinase-3, Noggin
Project description:The tightly controlled BMP-Smad1 pathway is essential for embryonic development and postnatal tissue homeostasis. Dysfunction of BMP-Smad1 signaling also leads to tumor development such as juvenile polyposis and Cowden syndromes and various tumors in mouse models, with unknown pathological mechanisms. Here we establish a link between the BMP-Smad pathway and the prominent tumor suppressor Atm-p53 pathway. We identify activated nuclear Smad1 as an Atm substrate under genotoxic stress. Atm-mediated Smad1 S239 phosphorylation disrupts Smad1 interaction with protein phosphatase PPM1A and enhances Smad1 activation and up-regulation, which not only turns on target genes including Cdk1nc but also interacts with p53 and inhibits Mdm2-mediated p53 ubiquitination, leading to p53 stabilization. Functionally, Smad1 acts like a tumor suppressor in DNA damage response, cell transformation and tumorigenesis in a p53-dependent manner. Sequencing of the gastric cancer samples revealed that Smad1 is frequently mutated, with S239 as mutational hotspot. This study thus establishes the BMP-Smad1 pathway as an integral part of DNA damage response, which can suppresses tumorigenesis via p53.