|
| Status |
Public on Apr 17, 2008 |
| Title |
Gene expression after infection of mice with Y. enterocolitica deltaYopH, lethal dose |
| Sample type |
RNA |
| |
|
| Source name |
CD11b+ cells of the spleen, infected of mice with Y. enterocolitica deltaYopH, lethal dose
|
| Organism |
Mus musculus |
| Characteristics |
Age: 6-8 weeks old
Gender: female
Strain: C57BL/6 mice
infected with 500000 Y. enterocolitica YopH deletion mutant (delta YopH), lethal dose
|
| Treatment protocol |
Mice were infected intravenously with 5000000 Y. enterocolitica YopH deletion mutant (DeltaYopH),lethal dose, for one day. After one day, five to ten mice per group were sacrificed by CO2 asphyxiation. The spleens were removed and placed in ice-cold HBSS (Ca2+ and Mg2+ free Hanks’ balanced salt solution [HBSS; BioWhittaker,Walkersville, MD.] supplemented with 2% FCS [HyClone, Logan, Utah] and 10 mM HEPES buffer). The spleens were forced with a 5 ml syringe pestle through a 100 µm-pore nylon mesh cell strainer (Falcon; BD Biosciences). Red blood cells were lysed from spleen samples by incubating the cell suspensions for 5 min at room temperature in erythrocyte lysis buffer (170 mM Tris, 160 mM NH4Cl, pH 7,4) followed by two washes in ice-cold HBSS. CD11b+ cells were purified by positive selection using MACS® CD11b MicroBeads (Miltenyi Biotec) according to the manufacturer´s instructions. To ensure enrichment values of 96-98% purity, the magnetic separation was performed twice. Cell viability and number were assessed by trypan blue exclusion.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was extracted from CD11b+ cells using the RNAeasy mini-kit following the manufacturer´s instructions (Qiagen, Hilden, Germany). Generation of fragmented cRNA was performed as recently described [38] and used for hybridization onto MG-U74Av2 from Affymetrix (Affymetrix, High Wycombe, UK). Genechips were washed, stained with streptavidin–phycoerythrin and read using a GeneChip Scanner 2500 (Affymetrix) at the IZKF microarray facility, Tübingen.
|
| Label |
Biotin
|
| Label protocol |
Standard procedure; Affymetrix protocol
|
| |
|
| Hybridization protocol |
Standard procedure; Affymetrix protocol
|
| Scan protocol |
Standard procedure; Affymetrix protocol
|
| Description |
CD11b+ cells of infected mice; Mice were infected with a lethal dose of Yersinia enterocolitica YopH deletion mutant(delta YopH) for one day
|
| Data processing |
Affymetrix ExpressionStat 5.0
|
| |
|
| Submission date |
Apr 16, 2008 |
| Last update date |
Apr 16, 2008 |
| Contact name |
Erwin Bohn |
| E-mail(s) |
[email protected]
|
| Phone |
00-49-70712981525
|
| Fax |
00-49-7071294972
|
| Organization name |
University of Tuebingen
|
| Department |
Institutefor Medical Microbiology and Hygiene
|
| Street address |
Elfriede-Aulhorn-Strasse 6
|
| City |
Tuebingen |
| ZIP/Postal code |
72076 |
| Country |
Germany |
| |
|
| Platform ID |
GPL81 |
| Series (1) |
| GSE11189 |
IFN-g counteracts YopH mediated immune evasion in Yersinia enterocolitica infection in mice |
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