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Sample GSM4512350 Query DataSets for GSM4512350
Status Public on Apr 12, 2021
Title 5 Gy control 3
Sample type SRA
 
Source name bone marrow endothelial cells
Organism Mus musculus
Characteristics strain: C57BL/6
radiation dose: 5 Gy
iv treatment: 10 ug IgG
Treatment protocol Mice were irradiated and treated daily for 2 days with with intravenous IgG control or anti-NRP1 antibody. At 72 hours, mice were euthanized and bone marrow endothelial cells were FACs sorted for ve-cadherin+cd45- cells.
Extracted molecule total RNA
Extraction protocol total RNA was isolated using the Qiagen microRNeasy kit
Libraries for RNA-Seq were constructed with Clontech Kit to generate strand-specific RNA-seq libraries. The workflow consists of poly(A) RNA selection, RNA fragmentation and double-stranded cDNA generation using a mixture of random and oligo(dT) priming, followed by end repair to generate blunt ends, adaptor ligation, strand selection, and PCR amplification to produce the final libraries
Amplified libraries were quantified by Qubit dsDNA HS (High Sensitivity) Assay Kit, and quality-checked by the Agilent 4200 TapeStation System. Different index adaptors were used for multiplexing samples in one sequencing lane. Sequencing was performed with NextSeq500 High Output sequencer to produce 75 base-pair single-end reads (1 x 75 bp).
 
Library strategy RNA-Seq
Library source transcriptomic
Library selection cDNA
Instrument model Illumina NextSeq 500
 
Description C3
Normalized_Gene_counts.txt
Data processing Reads per gene were quantified using STAR - 2.7.2a [3] and mm10 (Ensembl GRCm38.97). After obtaining gene counts, the counts were normalized by CPM. The Principal Component Analysis (PCA) was applied to the transcript counts. The differential gene expressions were examined by GSA. For all results of differential gene expression analysis, statistical filter were applied, which was p<0.05, FDR<0.05, and Fold Change |FC|>2.
Partek® Flow® software, version 7.0 Copyright ©; 2019 Partek Inc., St. Louis, MO, USA.
STAR: ultrafast universal RNA-seq aligner, A Dobin, CA Davis, F Schlesinger, J Drenkow, C Zaleski, S Jha, P Batut, Bioinformatics 29 (1), 15-21Fast gapped-read alignment with Bowtie 2. Langmead B, Salzberg SL. Nat Methods. (2012)
 
Submission date May 03, 2020
Last update date Apr 12, 2021
Contact name John P Chute
E-mail(s) John.Chute@cshs.org
Organization name University of California, Los Angeles (UCLA)
Street address 615 Charles E Young Drive South OHRC 545
City Los Angeles
State/province CA
ZIP/Postal code 90095
Country USA
 
Platform ID GPL19057
Series (1)
GSE149776 Bone Marrow Endothelial Cell Response to anti-NRP1 infusion
Relations
BioSample SAMN14823965
SRA SRX8244493

Supplementary data files not provided
SRA Run SelectorHelp
Raw data are available in SRA
Processed data are available on Series record

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