|
| Status |
Public on Dec 01, 2014 |
| Title |
129-Male Macrophages |
| Sample type |
RNA |
| |
|
| Source name |
Peritoneal macrophages isolated from 129S6/SvEvTac male mice
|
| Organism |
Mus musculus |
| Characteristics |
gender: male
strain: 129S6/SvEvTac
tissue: peritoneal macrophage
|
| Treatment protocol |
Three mice per group (malesand 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 C57BL/6 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 Affymetrix arrays. The Affymetrix HT WT User Manual was followed to prepare the samples. Briefly, the WT Expression HT Kit for Robotics (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 HT Terminal Labeling Kit.
|
| |
|
| Hybridization protocol |
The Beckman Coulter Biomek FXP Laboratory Automation Workstation with the Target Express set up was used to prepare the samples with these two kits. Fragmented and labeled cDNA was used to prepare a hybridization cocktail with the Affymetrix GeneTitan Hybridization Wash and Stain Kit for WT Arrays. Hybridization, washing, staining.
|
| Scan protocol |
Scanning of the Affymetrix array plate was carried out using the Affymetrix GeneTitan MC Instrument.
|
| 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 27, 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 |
GPL17400 |
| Series (2) |
| GSE52816 |
Expression data from thioglycollate-elicited peritoneal macrophages from 129S6/SvEvTac and C57BL/6 mice |
| GSE53006 |
Athero-susceptibility of inbred mouse strains |
|
