|
| Status |
Public on Dec 01, 2014 |
| Title |
DBA-Female Macrophages |
| Sample type |
RNA |
| |
|
| Source name |
Peritoneal macrophages isolated from DBA/2J female mice
|
| Organism |
Mus musculus |
| Characteristics |
gender: female
strain: DBA/2J
cell type: peritoneal macrophage
|
| Treatment protocol |
Three mice per group (males and females of each strain) were treated with 1 ml 4 % thioglycolate medium by intra-peritoneal injection and sacrificed 4 days after injection followed by intra-peritoneal lavage with 10 ml PBS. Macrophages were plated at in 100 mm plates and cultured overnight in DMEM supplemented with 10% FBS.
|
| Growth protocol |
Wild type 129S6/SvEvTac and DBA2/J mice mice were fed with a regular chow.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was extracted by RNeasy kit (Qiagen) according to the manufacturer's instructions. Equal amounts of RNA from three mice in each group were pooled.
|
| Label |
biotin
|
| Label protocol |
Total RNA (250 ng) was used to synthesize fragmented and labeled sense-strand cDNA and hybridize onto Affymetrx arrays. Briefly, the WT Expression Kit (Ambion) was used to generate sense-strand cDNA from total RNA. Following synthesis of sense-strand cDNA, the cDNA was fragmented and labeled with the Affymetrix GeneChip Terminal Labeling Kit. Fragmented and labeled cDNA was then added to a hybridization cocktail (25 µg/µl fragmented cDNA, 50 pM control oligonucleotide B2, BioB, BioC, BioD and cre hybridization controls, 7 % DMSO, 100 mM MES, 1 M [Na+], 20 mM EDTA, 0.01% Tween 20).
|
| |
|
| Hybridization protocol |
Affymetrix arrays (Affymetrix, Santa Clara, CA) were hybridized for 16 hours at 45°C in the GeneChip Hybridization Oven 640 (Affymetrix). The arrays were washed and stained with R-phycoerythrin streptavidin in the GeneChip Fluidics Station 450 (Affymetrix).
|
| Scan protocol |
The arrays were scanned with the GeneChip Scanner 3000 7G Plus with autoloader. GeneChip Command Console Software (AGCC) was used for washing, staining and scanning control of the instrumentation.
|
| Description |
Gene expression data of peritoneal macrophages pooled from three mice.
|
| Data processing |
Affymetrix Expression Console Software was used for basic data analysis and quality control.
|
| |
|
| Submission date |
Nov 18, 2013 |
| Last update date |
Dec 01, 2014 |
| Contact name |
Yukako Kayashima |
| E-mail(s) |
yukaya@email.unc.edu
|
| Organization name |
University of North Carolina at Chapel Hill
|
| Street address |
701 Brinkhous-Bullitt Building
|
| City |
Chapel Hill |
| State/province |
NC |
| ZIP/Postal code |
27599 |
| Country |
USA |
| |
|
| Platform ID |
GPL16570 |
| Series (2) |
| GSE52473 |
Expression data from thioglycollate-elicited peritoneal macrophages from 129S6/SvEvTac and DBA2/J mice |
| GSE53006 |
Athero-susceptibility of inbred mouse strains |
|
