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Parallel single-cell bisulfite- and RNA-sequencing link transcriptional and epigenetic heterogeneity [RNA-seq]

ABSTRACT: We report scM&T-seq, a method for parallel single-cell genome-wide methylome and transcriptome sequencing, allowing discovery of associations between transcriptional and epigenetic variation. Profiling of 61 mouse embryonic stem cells confirmed known links between DNA methylation and transcription. Notably, the method reveals novel associations between heterogeneous methylation of distal regulatory elements and transcriptional heterogeneity of key pluripotency genes. E14 ES cells were grown in either serum/LIF or 2i culture conditions and separated into single cells. RNA-Seq or Bisulfite-Seq libraries were prepared. This Series includes only the RNA-Seq data. The list of 61 samples that passed QC in both BS-seq and RNA-seq is included in "Supplementary Table 1" of the associated manuscript.

ORGANISM(S): Mus musculus

SUBMITTER: Felix Krueger

PROVIDER: E-GEOD-74534 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Parallel single-cell bisulfite- and RNA-sequencing link transcriptional and epigenetic heterogeneity [Bisulfite-Seq]

Project description:We report scM&T-seq, a method for parallel single-cell genome-wide methylome and transcriptome sequencing, allowing discovery of associations between transcriptional and epigenetic variation. Profiling of 61 mouse embryonic stem cells confirmed known links between DNA methylation and transcription. Notably, the method reveals novel associations between heterogeneous methylation of distal regulatory elements and transcriptional heterogeneity of key pluripotency genes. E14 ES cells were grown in either serum/LIF or 2i culture conditions and separated into single cells. RNA-Seq or Bisulfite-Seq libraries were prepared. This Series includes only the Bisulfite-Seq data. The list of 61 samples that passed QC in both BS-seq and RNA-seq is included in "Supplementary Table 1" of the associated manuscript.

2015-12-24 | E-GEOD-68642 | biostudies-arrayexpress

Parallel single-cell bisulfite- and RNA-sequencing link transcriptional and epigenetic heterogeneity

Project description:This SuperSeries is composed of the SubSeries listed below. Refer to individual Series

2015-12-24 | E-GEOD-74535 | biostudies-arrayexpress

Parallel single-cell bisulfite- and RNA-sequencing link transcriptional and epigenetic heterogeneity [RNA-seq]

2015-12-24 | GSE74534 | GEO

Parallel single-cell bisulfite- and RNA-sequencing link transcriptional and epigenetic heterogeneity [Bisulfite-Seq]

2015-12-24 | GSE68642 | GEO

Sex comb on midleg (Scm) is a functional link between PcG repressive complexes in Drosophila

Project description:We profiled Sex comb on midleg (Scm), Pc and E(z) in fly embryos and S2 based on BioTAP-XL ChIP-seq. ChIP-seq revealed that Scm is co-localized with PRC1, PRC2, and H3K27me3 in both S2 cells and embryos. Genomic binding/occupancy profiling of Scm, Pc and E(z) by high throughput sequencing

2015-07-09 | E-GEOD-66183 | biostudies-arrayexpress

Sex comb on midleg (Scm) is a functional link between PcG repressive complexes in Drosophila

2015-07-09 | GSE66183 | GEO

Transcriptomics Changes of Calcitonin Gene-related Peptide in Mitigating Lipopolysaccharide-induced Septic Cardiomyopathy

Project description:Septic cardiomyopathy (SCM) is the predominant cause of death in sepsis patients with undefined mechanism to date. Our study analyzed RNA sequencing (RNA-seq) data from rat heart tissue to discover key targets and potential pharmacological actions of the calcitonin gene-related peptide (CGRP) against SCM. A lipopolysaccharide-induced SCM model was established in rats (LPS 10 mg/kg, intraperitoneal (i.p.)). Thereafter, the myocardial tissues from the three groups of rats (Ctrl group, LPS group, and CGRP group) (n=5) were extracted and underwent RNA-seq, followed by bioinformatics analyses. Our findings suggest a basis for finding potential targets for CGRP in the treatment of SCM.

2024-11-06 | GSE266124 | GEO

Enrichment and quantitation of newly secreted proteins under serum condition by combining BONCAT and pulsed SILAC

Project description:Despite the increasing interest of secretome involved in paracrine/autocrine mechanisms, the majority of the mass spectrometric cell secretome studies have been performed using serum-free medium (SFM), due to low-abundance of secreted proteins and high-abundance of contaminating fetal bovine serum (FBS) proteins in serum-containing medium (SCM). In this study, through the combination of bioorthogonal non-canonical amino acid tagging (BONCAT) and pulsed-SILAC (pSILAC), the low-abundant secreted proteins were captured selectively and quantified. We set up the combined BONCAT-pSILAC method by optimizing capturing condition and by using a composite human-FBS database, and applied it to the analysis of differentially secreted proteins between SFM and SCM. The amount of secreted proteins for 24 h in SCM was much bigger than that from SFM for U-87MG glioblastoma cells (1.28-fold) and mesenchymal stem cells from human umbilical cord blood (hUCB-MSCs; 2.01-fold). The proteins secreted more in SCM were predominantly predicted to be truly secretory. Therefore, this study suggests the analysis of the secretome should be processed in serum-containing medium that promotes cell proliferation and secretion.

2019-03-13 | PXD007529 | Pride

Proteomic Comparison of Acute Myocardial Infarction and Stress Cardiomyopathy in Women

Project description:Background: Stress cardiomyopathy (SCM) is a unique form of LV dysfunction that more often occurs in women. Patients with SCM have a higher Troponin I/B-type natriuretic peptide ratio than AMI, but little is known about other circulating proteins. The goals of this study were to compare plasma proteins in SCM and AMI to learn about the pathophysiology of SCM and also to identify putative biomarkers of SCM. Methods: Blood was drawn in normal controls (n=6), women with AMI (n=12) or women with acute SCM (n=15). Two-week follow up samples were available in AMI (n=4) and SCM patients (n=11). Relative concentrations of 1310 serum proteins were measured in each of the 48 samples using the SOMAscan aptamer based assay. Women with AMI tended to be younger (57.87 ± 16.0 vs. 65.08 ± 9.11 years, p=0.12) and had a higher peak troponin I (AMI 32.03 ± 29.46 vs. SCM 2.68 ± 2.6 ng/mL, p=0.02). No differentially expressed proteins were detected (absolute log2 fold change>1; q<0.05) between AMI and SCM in the acute or recovery phase. In the normal vs. AMI comparison there was differential expression of 35 proteins. In the normal vs. SCM comparison there were 45 proteins with differential expression. Pathway analysis demonstrated activation of complement, coagulation, and inflammation in both AMI and SCM. Conclusions: The acute phase of SCM is characterized by a severe inflammatory response similar to AMI. Despite recovery of LV function in SCM at two weeks, differences in circulating proteins remain in comparison to normal controls.

2017-08-03 | GSE95368 | GEO

Limitations and possibilities of low cell number ChIP-seq

Project description:ChIP-seq experiments using low numbers of input cells, scaled down to the point where data quality is unnacceptably compromised, reveals limits of the technique. Two-part ChIPseq study using native chromatin (non-crosslinked) generated with MNase from human CD4+ cells. Part 1: Previously published protocol (Barski et al, 2007, Cell 129:823, hereafter called "Benchmark") used with 2x10e7 cells / ChIP with H3K4me3 antibody, compared to modified method ("new") with decreasing input cell numbers spanning 1000-fold range from 2x10e7 cells / IP to 2x10e4 cells/ IP. Part 2: reproducibility of new method studied using triplicate samples at lowest two cell numbers tested: 1x10e5 and 2x10e4 / ChIP, using H3K4me3 and H3K27me3 antisera.

2012-12-03 | E-GEOD-42147 | biostudies-arrayexpress

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