|
| Status |
Public on May 30, 2014 |
| Title |
15hControl1 |
| Sample type |
RNA |
| |
|
| Source name |
GEO_15C1
|
| Organism |
Sus scrofa |
| Characteristics |
cell type: porcine alveolar macrophage
infection: none
time point: 15 hours
|
| Treatment protocol |
Cells and tracheal explants from each of the donor pigs were subjected to all treatments. M. hyopneumoniae was inoculated onto the tracheal rings of the appropriate groups 6 h prior to their addition to the PAMs. PAMs from the PRRSV-infected groups were inoculated with PRRSV 1 h prior to the addition of the tracheal rings. The tracheal ring inserts and their supernatants were transferred to the cultured PAMs, and the mixture was incubated at 37°C in 5%CO2 for 6 or 15 hpi prior to harvesting of the PAMs with a cell scraper. The supernatant from the tracheal cultures was dripped through the insert over the PAMs so no cell-to-cell contact between the tracheal rings and PAMs occurred
|
| Growth protocol |
Porcine alveolar macrophages were obtained with consent from one healthy pig (serologically negative for PRRSV, M. hyopneumoniae and porcine circovirus type 2)
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Trizol extraction of total RNA was performed according to the manufacturer's instructions.
|
| Label |
Cy3
|
| Label protocol |
5 mg cRNA was reverse transcribed to cDNA, fragmented, and then labeled with Cy3-dCTP (GE Healthcare) using Klenow.
|
| |
|
| Hybridization protocol |
These labeled cDNA fragments were hybridized to NimbleGen Porcine Genome Expression Arrays for 16 h at 42uC using the Roche NimbleGen Hybridization System 12.
|
| Scan protocol |
Array scanning using the Axon GenePix 4000B microarray scanner (Molecular Devices Corporation).
|
| Description |
uninfection
|
| Data processing |
Scanned images (TIFF format) were then imported into NimbleScan software (version 2.5) for grid alignment and expression data analysis.Expression data were normalized through quantile normalization and the Robust Multichip Average (RMA) algorithm included in the NimbleScan software.
|
| |
|
| Submission date |
Aug 14, 2013 |
| Last update date |
May 30, 2014 |
| Contact name |
li bin |
| E-mail(s) |
libinana@126.com
|
| Phone |
86-25-84390330
|
| Organization name |
Jiangsu Academy of Agricultural Sciences
|
| Street address |
Zhong-ling street 50
|
| City |
Nanjing |
| State/province |
Jiangsu |
| ZIP/Postal code |
210014 |
| Country |
China |
| |
|
| Platform ID |
GPL17577 |
| Series (1) |
| GSE49882 |
Transcription analysis of the porcine alveolar macrophage （PAM） response to co-infection of porcine reproductive and respiratory syndrome virus (PRRSV) and M. hyopneumoniae (Mhp) |
|
