RNA-seq profiling of FACS purified MPs from adult skeletal muscle after conditional knockout of Hic1 [MP RNA-seq]
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ABSTRACT:
PROVIDER: PRJNA432558 | ENA |
REPOSITORIES: ENA
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Project description:To determine the effect of the deletion of the trancriptional repressor Hypermethylated in cancer 1 (Hic1) on quiescence, adult mesenchymal progenitors (MPs) were isolated from skeletal muscle of both wild type mice and those conditionally deleted for Hic1 and profiled by RNA sequencing. Comparison to MPs activated by using a muscle injury model of the tibialis anterior muscle revealed similar pathways affected under both conditions, highlighting the role for Hic1 in maintaining the quiescent state of MPs
2019-12-05 | GSE110012 | GEO
Project description:To determine the effects of long term deletion of the transcriptional repressor Hypermethylated in cancer 1 (Hic1) on skeletal muscle, samples were collected of both wild type mice and those conditionally deleted for Hic1 and profiled by RNA sequencing. Aspects of muscle homeostasis were negatively impacted, highlighting cell and non cell autonomous roles for Hic1 in maintaining functional MPs and muscle tissue.
2019-12-05 | GSE133995 | GEO
Project description:The important role of mesenchymal progenitors is quickly emerging in various biological processes. Thus far an appropriate marker has been lacking. Here the fractionation of dissociated skeletal muscle and subsequent enrichment of MPs by an established method was employed. Endothelial and blood derived cells comprised the lineage positive fraction and the remaining cell suspension was fractionated using the surface marker Sca1(Ly6a) to enrich for MPs. RNA sequencing was performed on all fractions and subsequent transcriptomic data was interrogated for MP specific transcription factors. Hic1 was found to be highly enriched in the MP fraction.
2019-12-05 | GSE110013 | GEO
Project description:To further elucidate the mechanism underlying the RT-MPs killing effect on SLTCs, we performed RNA-seq on stem-like Lewis cells and RT-MP-treated stem-like Lewis cells We then performed gene expression profiling analysis using data obtained from RNA-seq of stem-like Lewis cells untreated or treated with RT-MPs for 48 h
2023-05-30 | GSE233577 | GEO
Project description:3 subtypes of cortical projection neurons were purified by fluorescence-activated cell sorting at 4 different stages of development from mouse cortex. A detailed description of the data set is described in Arlotta, P et al (2005). Keywords = corticospinal motor neuron callosal corticotectal cortex development FACS
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Project description:Background Proteomic characterization of microglia has been limited by low yield and contamination by non-microglial proteins by magnetic-activated cell sorting (MACS) enrichment strategies. To determine whether a fluorescent-activated cell sorting (FACS)-based strategy overcomes these limitations, we compared microglial proteomes of MACS and FACS-based purified CD11b+ microglia in order to identify core sets of microglial proteins in adult mouse brain tissue. Results Quantitative multiplex proteomics by tandem mass tag mass spectrometry (TMT-MS) of MACS-enriched (N = 5) and FACS-purified (N = 5) adult wild-type CD11b+ microglia identified 1,791 proteins, of which 953 were differentially expressed indicating significant differences between both approaches. While the FACS-purified microglia proteome was enriched with cytosolic, endoplasmic reticulum and ribosomal proteins involved in protein metabolism and immune system functions, the MACS-enriched microglia proteome was enriched with proteins related to mitochondrial function and synaptic transmission. As compared to MACS, the FACS microglial proteome showed strong enrichment for canonical microglial proteins while neuron, astrocyte, and oligodendrocyte proteins were decreased. Interestingly, we observed enrichment of endothelial specific proteins in the FACS microglia proteome. By comparing FACS-purified microglia proteomes with transcriptomes, we observed highly concordant as well as highly discordant proteins that were abundant at the protein level but low at the transcript level. Conclusions We demonstrate that TMT-MS proteomics of FACS-purified adult microglia is superior to column-based enrichment approaches, resulting in purer and highly-enriched microglial proteomes. We also define core sets of highly-abundant adult microglial proteins including Moesin (Msn) that can guide future studies.
2020-05-11 | PXD015652 | Pride
Project description:Background-The endothelial protein C receptor (EPCR) plays an important role within the protein C (PC) pathway in regulating coagulation and inflammation. Recently, we described a novel mechanism of EPCR release from the surface of primary physiological cells. Induced by exogenous activated protein C (APC), EPCR is released in microparticulate form. Its bound APC retains proteolytic anticoagulant activity and we now hypothesise that this microparticulate EPCR-APC complex can also cleave endothelial protease activated receptor 1 (PAR1) to modulate inflammation and cytoprotection. Methods & Results-The gene profiling effect on human endothelial cells by 40nM APC, in free or microparticulate form, was assessed. Transcript profile results showed upregulation of anti-apoptotic and inflammatory genes by APC in either form, which were confirmed by RT-PCR. These translated into increased GM-CSF and interleukin 8 secretion and cytoprotection against staurosporine-induced apoptosis. PC or PAR1 antagonism reversed these results to demonstrate that induced effects by microparticles were APC specific and PAR1-dependent. Further analysis of human plasma from septic patients, by confocal microscopy and ELISA, showed evidence of circulating microparticle-associated EPCR during recombinant APC treatment. Functional coagulation and cellular studies demonstrate that these express APC-specific effects on anticoagulation with PAR1-mediated gene induction and anti-apoptotic function. Conclusions-APC induction of microparticle-associated EPCR release can occur in vivo. These microparticles could potentially disseminate the function of the EPCR-APC complex to PAR1 on different cells and vascular sites. Keywords: pre versus rhAPC-treatment mp in vivo MP were isolated from sepsis patients pre and during-rhAPC treatment. MP were counted by FACS using CD13 positivity and same number of mp was used in pre and during treatment comparisons. HUVEC were incubated with the in vivo mps according to mp number or apc content, and controls for both were done using non-treated patients or free-apc. The role of apc and par1 was analysed by including apc blocking antibody or par1 antaginist peptide respectively.
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Project description:We sequenced mRNA from HFSCs that were FACS purified (CD34+alpha-6 Integrin+ cells) from p21 mouse epidermis, total epidermal cells from p21 mice and HFSCs cultured for 14 days in 3C cultures, 3 biological replicates each. We sequenced mRNA from different cell populations that were FACS-purified from HFSC cultures (also known as 3C cultures), 3 biological replicates each. We performed two independent comparisons: 1. Comparison of FACS-purified cells grown in standard 3C cultures: HFSCs (CD34+ alpha-6 Integrin+) and non-HFSCs (CD34neg alpha-6 Integrin+). 2. Comparison of total cells in 3C cultures treated for 48 hours with dorsomorphin (BMP inhibitor that causes depletion of CD34+ alpha-6 integrin+ cells in 3C cultures) or with cyclopamine Shh inhibitor that causes enrichment of CD34+ alpha-6 integrin+ cells in 3C cultures).
2016-12-09 | GSE76779 | GEO
Project description:The tumor suppressor gene HIC1 (Hypermethylated in Cancer 1) is essential for mammalian development and is epigenetically silenced in many human tumors. Functionally, HIC1 is involved in a complex pathway regulating P53 tumor-suppression activity. HIC1 encodes a sequence-specific transcriptional repressor containing five Krüppel-like C2H2 zinc fingers but only a few genes regulated by HIC1 have been reported, including SIRT1. Keywords: Time Series Design
2007-12-13 | GSE9854 | GEO
Project description:This SuperSeries is composed of the following subset Series:; GSE2039: FACS purified cortical projection neurons; GSE17783: Analysis of gene expression in FACS-purified cortical projection neurons using Affymetrix 430 2.0 microarrays Experiment Overall Design: Refer to individual Series
2009-10-10 | E-GEOD-17784 | biostudies-arrayexpress