2114f454-2241-4480-9997-8e157413fa77 - samples
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ABSTRACT:
PROVIDER: EGAD00001007024 | EGA |
REPOSITORIES: EGA
Similar Datasets
Project description:The dataset includes RNA-seq expression R data, RNA-seq gene counts matrix, and RNA-seq gene FPKM matrix from 21 NEN tumors or metastases and matched PDTOs. The sequencing was done with either a Nextseq 2000 or a NovaSeq 6000 instrument.
| EGAD00001009994 | EGA
Project description:In an effort to confirm binding of SOMAmer®s to their respective targets in an endogenous matrix, we have conducted a series of experiments using SOMAmer®s to enrich target proteins followed by measurement using two mass spectrometry techniques: data dependent analysis (DDA) and multiple reaction monitoring (MRM). For data dependent analysis, the library of 4783 SOMAmer®s were multiplexed in sets of 8 and used for enrichment of target proteins from cell lysate and conditioned media, as well as human plasma and serum. A subset of the SOMAmer®s were screened in urine by this global profiling technique as well. Cell lines were selected from Cancer Cell Line Encyclopedia for screening by comparing gene expression of the target proteins measured by RNA sequencing across the CCLE.1 Using the criterion of FPKM value greater than 5 for the target protein, the minimum number of cell lines was grown to maximize protein coverage across the SOMAmer® library. For a given cell line, the presence of at least 8 target proteins with FPKM values greater than 5was required for inclusion. Approximately 400 target proteins were not covered by this strategy were screened in serum and plasma only. For enrichment from biological matrices, SOMAmer®s were combined into sets of 8 such that the potential interaction with similar proteins or binding partners was minimized. Non-specific binding of the SOMAmer®s was assessed in each matrix by using a SOMAmer® generated against the bacterial protein CysH. Target protein spectral counts in the SOMAmer® enriched samples were compared to the respective CysH control. A positive hit is defined as target protein detection with a minimum of 2 spectral counts and signal over CysH background greater than 10X (if applicable).
2018-08-03 | PXD008820 | Pride
Project description:In an effort to confirm binding of SOMAmer®s to their respective targets in an endogenous matrix, we have conducted a series of experiments using SOMAmer®s to enrich target proteins followed by measurement using two mass spectrometry techniques: data dependent analysis (DDA) and multiple reaction monitoring (MRM). For data dependent analysis, the library of 4783 SOMAmer®s were multiplexed in sets of 8 and used for enrichment of target proteins from cell lysate and conditioned media, as well as human plasma and serum. A subset of the SOMAmer®s were screened in urine by this global profiling technique as well. Cell lines were selected from Cancer Cell Line Encyclopedia for screening by comparing gene expression of the target proteins measured by RNA sequencing across the CCLE.1 Using the criterion of FPKM value greater than 5 for the target protein, the minimum number of cell lines was grown to maximize protein coverage across the SOMAmer® library. For a given cell line, the presence of at least 8 target proteins with FPKM values greater than 5was required for inclusion. Approximately 400 target proteins were not covered by this strategy were screened in serum and plasma only. For enrichment from biological matrices, SOMAmer®s were combined into sets of 8 such that the potential interaction with similar proteins or binding partners was minimized. Non-specific binding of the SOMAmer®s was assessed in each matrix by using a SOMAmer® generated against the bacterial protein CysH. Target protein spectral counts in the SOMAmer® enriched samples were compared to the respective CysH control. A positive hit is defined as target protein detection with a minimum of 2 spectral counts and signal over CysH background greater than 10X (if applicable).
2018-08-03 | PXD008819 | Pride
Project description:In an effort to confirm binding of SOMAmer®s to their respective targets in an endogenous matrix, we have conducted a series of experiments using SOMAmer®s to enrich target proteins followed by measurement using two mass spectrometry techniques: data dependent analysis (DDA) and multiple reaction monitoring (MRM). For data dependent analysis, the library of 4783 SOMAmer®s were multiplexed in sets of 8 and used for enrichment of target proteins from cell lysate and conditioned media, as well as human plasma and serum. A subset of the SOMAmer®s were screened in urine by this global profiling technique as well. Cell lines were selected from Cancer Cell Line Encyclopedia for screening by comparing gene expression of the target proteins measured by RNA sequencing across the CCLE.1 Using the criterion of FPKM value greater than 5 for the target protein, the minimum number of cell lines was grown to maximize protein coverage across the SOMAmer® library. For a given cell line, the presence of at least 8 target proteins with FPKM values greater than 5was required for inclusion. Approximately 400 target proteins were not covered by this strategy were screened in serum and plasma only. For enrichment from biological matrices, SOMAmer®s were combined into sets of 8 such that the potential interaction with similar proteins or binding partners was minimized. Non-specific binding of the SOMAmer®s was assessed in each matrix by using a SOMAmer® generated against the bacterial protein CysH. Target protein spectral counts in the SOMAmer® enriched samples were compared to the respective CysH control. A positive hit is defined as target protein detection with a minimum of 2 spectral counts and signal over CysH background greater than 10X (if applicable).
2018-08-03 | PXD008821 | Pride
Project description:In an effort to confirm binding of SOMAmer®s to their respective targets in an endogenous matrix, we have conducted a series of experiments using SOMAmer®s to enrich target proteins followed by measurement using two mass spectrometry techniques: data dependent analysis (DDA) and multiple reaction monitoring (MRM). For data dependent analysis, the library of 4783 SOMAmer®s were multiplexed in sets of 8 and used for enrichment of target proteins from cell lysate and conditioned media, as well as human plasma and serum. A subset of the SOMAmer®s were screened in urine by this global profiling technique as well. Cell lines were selected from Cancer Cell Line Encyclopedia for screening by comparing gene expression of the target proteins measured by RNA sequencing across the CCLE.1 Using the criterion of FPKM value greater than 5 for the target protein, the minimum number of cell lines was grown to maximize protein coverage across the SOMAmer® library. For a given cell line, the presence of at least 8 target proteins with FPKM values greater than 5was required for inclusion. Approximately 400 target proteins were not covered by this strategy were screened in serum and plasma only. For enrichment from biological matrices, SOMAmer®s were combined into sets of 8 such that the potential interaction with similar proteins or binding partners was minimized. Non-specific binding of the SOMAmer®s was assessed in each matrix by using a SOMAmer® generated against the bacterial protein CysH. Target protein spectral counts in the SOMAmer® enriched samples were compared to the respective CysH control. A positive hit is defined as target protein detection with a minimum of 2 spectral counts and signal over CysH background greater than 10X (if applicable).
2018-08-03 | PXD008823 | Pride
Project description:In an effort to confirm binding of SOMAmer®s to their respective targets in an endogenous matrix, we have conducted a series of experiments using SOMAmer®s to enrich target proteins followed by measurement using two mass spectrometry techniques: data dependent analysis (DDA) and multiple reaction monitoring (MRM). For data dependent analysis, the library of 4783 SOMAmer®s were multiplexed in sets of 8 and used for enrichment of target proteins from cell lysate and conditioned media, as well as human plasma and serum. A subset of the SOMAmer®s were screened in urine by this global profiling technique as well. Cell lines were selected from Cancer Cell Line Encyclopedia for screening by comparing gene expression of the target proteins measured by RNA sequencing across the CCLE.1 Using the criterion of FPKM value greater than 5 for the target protein, the minimum number of cell lines was grown to maximize protein coverage across the SOMAmer® library. For a given cell line, the presence of at least 8 target proteins with FPKM values greater than 5was required for inclusion. Approximately 400 target proteins were not covered by this strategy were screened in serum and plasma only. For enrichment from biological matrices, SOMAmer®s were combined into sets of 8 such that the potential interaction with similar proteins or binding partners was minimized. Non-specific binding of the SOMAmer®s was assessed in each matrix by using a SOMAmer® generated against the bacterial protein CysH. Target protein spectral counts in the SOMAmer® enriched samples were compared to the respective CysH control. A positive hit is defined as target protein detection with a minimum of 2 spectral counts and signal over CysH background greater than 10X (if applicable).
2018-08-03 | PXD008822 | Pride
Project description:Primary outcome(s): Cancer-related gene alteration, expression of protein, and expression of RNA
| 2645675 | ecrin-mdr-crc
Project description:Histone 3 lysine 4 trimethylation (H3K4me3) is known to be associated with transcriptionally active or poised genes and required for postnatal neurogenesis within the subventricular zone (SVZ) in the rodent model. Previous comparisons have shown significant correlation between baboon (Papio anubis) and human brain. In this study, we demonstrate that chromatin activation mark H3K4me3 is present in undifferentiated progenitor cells within the SVZ of adult baboon brain. To identify the targets and regulatory role of H3K4me3 within the baboon SVZ, we developed a technique to purify undifferentiated SVZ cells while preserving the endogenous nature without introducing culture artifact to maintain the in vivo chromatin state for genome-wide studies (ChIP-Seq and RNA-Seq). We identified that H3K4me3 is significantly enriched for genes involved in cell cycle, cell signaling, nervous system development, metabolism, and ribosomal biogenesis. Among these genes, RNA-Seq identified that genes associated with cellular signaling/maintenance, DNA replication, metabolism, and protein synthesis are expressed. We found that 72% of H3K4me3-enriched genes in undifferentiated SVZ cells are expressed (1913 genes are detectable by RNA-Seq; 2663 genes enriched with H3K4me3 by ChIP-Seq). On the other hand, 750 out of total 2663 H3K4me3-enriched genes are not detectable by RNA-Seq, suggesting 28% of genes in SVZ are poised for activation. RNA-seq profiles were generated from adult baboon subventricular zone primary cells by paired-end deep sequencing with an Illumina HiSeq 2000. RNA-Seq Analysis: Total RNA was extracted from purified baboon SVZ cells using TRIzol reagent, and sequencing libraries were generated with the Illumina RNA-Seq library kit. Paired-end RNA-deepSeq (76 base pair; >300 million tag reads; 269,081,636 mapped reads) were aligned to hg19. DESeq was used to normalize raw read counts, and Cufflink reports read counts and estimated FPKM (fragments per kilobase of exon per million fragments mapped; http://cufflinks.cbcb.umd.edu/faq.html#fpkm). Genes with expression values >1 FPKM were considered for subsequent analyses.
2014-07-16 | E-GEOD-58527 | biostudies-arrayexpress
Project description:Total RNA of regulatory T (Treg) cells and conventional CD4 T (Tconv) cells from young WT (2 months), aged WT (>18 months) and young Dcaf1-deficient mice was extracted and RNA-seq libraries were generated. Reads (32-45 Million reads per sample) were analyzed with Salmon software to align and quantify the transcript expression. R packages in Bioconductor, tximport and tximportData were used to aggregate transcript-level quantifications to the gene level, with the R package biomaRt for gene and transcripts mapping. The option "lengthScaledTPM" for countsFromAbundance in tximport was used to obtain the estimated counts at the gene level using abundance estimates scaled based on the average transcript length over samples and the library size. The option "lengthScaledTPM" for countsFromAbundance in tximport was used to obtain the estimated counts at the gene level using abundance estimates scaled based on the average transcript length over samples and the library size. For the differential expression (DE) analysis of RNA-seq data, gene-level count matrix was passed into by DESeq2 package as input directly from the tximport package. The normalized gene expression data was retrieved from DESeq2 analysis after regulated log (rlog) transformation (‘rlog’ in DESeq2). The z-score at gene-level average of normalized expression matrix was used to generate heatmap in Gene-E from Broad Institute (www.broadinstitute.org/ /GENE-E/). Gene Set Enrichment Analysis was performed using the Java application available from Broad Institute (www.broadinstitute.org/gsea/). Gene set databases including Hallmarks (h.all.v6.1.symbols.gmt) and KEGG (c2.cp.kegg.v6.1.symbols.gmt) from the Molecular Signatures Database (MSigDB) were used in the analysis. The aging-program gene set was from DEMAGALHAES_AGING_UP in MSigDB. One thousand gene set permutations were performed. FDR<0.05 was used for enriched terms, as is recommended when performing permutations by gene set.
2020-10-10 | GSE130419 | GEO
Project description:The goals of this study are to comprehensively identify genes controlled by Jmjd6 in the thymic stroma, and to identify a novel alternative splicing mechanism. Methods: Samples were WT and Jmjd6-/- fetal thymus organ culture with (2 samples for each category) or without (1 sample for each category) RANKL stimulation for 4 days under 2-DG. One µg of total RNA was used for library construction with TruSeq RNA Sample Prep Kit v2. THe ligated products were amplified using 8 cycles of PCR to generate RNA-seq library. Library integrity was verified by Bioanalyzer DNA1000 assay. Sequencing was performed in 101-bp paired-end mode using an Illumina HiSeq.Technical duplicate has done. Results: A total of 177,060,020 reads were obtained for 6 samples. Filtered reads were mapped to the UCSC mm10 using the TopHat program(v2.0.10) with the default parameters. The Cufflinks program (v2.1.1) was then used to assemble 22,448 transcripts and to calculate the fragments per kilobase of exon per million mapped fragments(FPKM) values, which are normalized measurement of gene expression levels(= genes-FPKM file).To identify differentially expressed genes, the ratio of the maximum FPKM to the minimum FPKM was compared among 6 samples. When the ratio was more than 3, the gene was regarded as being significantly altered in expression level. We added 0.1 to the FPKM value to avoid division by zero. This led us to identify 3212 genes with differential expression. Among these, the expression levels of 2536 genes were significantly associated with the RANKL treatment or Jmjd6 expression ( P value <0.05). Analysis for intron retention was performed as follows. According to the current gene annotation ("known genes" in UCSC mm10), there are 188,208 introns in total. As intron retention events should be observed in the genes with relatively high expression, we only focused on the genes with the maximum FPKM value more than 10 at least in one of the six samples. As a result, we obtained 84,708 introns. The reads mapped to these intronic regions were counted by the intersectBed program in the BEDTools utilities with -c option, and the counts are converted into the FPKM values for each intron (intronic FPKM). There are 1051 introns with intronic FPKM more than 10 in at last on of the six samples, and the degree of intron retention was calculated by dividing intronic FPKM value by conventional FPKM value for each gene (intron-FPKM file). Samples were WT and Jmjd6-/- fetal thymus organ culture with (2 samples for each category) or without (1 sample for each category) RANKL stimulation for 4 days under 2-DG. One µg of total RNA was used for library construction with TruSeq RNA Sample Prep Kit v2. THe ligated products were amplified using 8 cycles of PCR to generate RNA-seq library. Library integrity was verified by Bioanalyzer DNA1000 assay. Sequencing was performed in 101-bp paired-end mode using an Illumina HiSeq.Technical duplicate has done.
2015-10-22 | E-GEOD-61444 | biostudies-arrayexpress