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| Status |
Public on Jun 08, 2012 |
| Title |
Comparison of gene expression in NOD versus NOD.NOR-Chr4 (NR4) splenic B cells. |
| Organism |
Mus musculus |
| Experiment type |
Expression profiling by array
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| Summary |
Type 1 Diabetes (T1D) in humans and the non-obese diabetic (NOD) mouse model results from autoreactive T cell destruction of pancreatic beta cells. A pathogenic role for B lymphocytes (B cells) in T1D first became evident when NOD mice made deficient in this population through introduction of an inactivated Igµ heavy chain gene (NOD.Igµnull) or chronic treatment with anti-IgM antibodies were strongly protected from disease. We produced an NOD background strain developing a greatly decreased T1D incidence due to a NOR-derived 44.31Mb congenic region from rs3674285 to D4Mit127 on distal Chr. 4 (termed NOD.NOR-Chr4 (NR4)) containing disease resistance alleles decreasing the pathogenic activity of autoreactive B cells. Microarrays were conducted on B cells purified from spleens of NOD and NR4 mice to highlight differentially expressed genes within the distal Chr. 4 locus. B cells were either cultured in media alone (unstimulated) or with BCR cross-linking anti-IgM-F(ab’)2 fragments (stimulated) for 2h before RNA was extracted for transcript analysis.
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| Overall design |
B cells from three independent lots of pooled spleens (n=7-8) from NOD or NR4 female mice, respectively, were purified using a magnetic-activated cell sorting (MACS) negative depletion strategy. Purified B cells from each lot were resuspended at 1x10<7> cells/ml for 2h in complete RPMI media alone or with 10µg/ml AffiniPure goat anti-mouse IgM F(ab’)2 fragments. RNA was then purified from B cells and subjected to one-cycle linear amplification and biotin labeling. Two BCR-stimulated and three unstimulated NOD B cell samples plus three BCR-stimulated and three unstimulated NR4 B cell samples were hybridized to Mouse Genome 430 2.0 Arrays.
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| Contributor(s) |
Silveira PA, Serreze DV |
| Citation(s) |
22732593 |
| Submission date |
Apr 16, 2012 |
| Last update date |
Feb 11, 2019 |
| Contact name |
Pablo Alejandro Silveira |
| E-mail(s) |
P.Silveira@garvan.org.au
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| Phone |
+61-2-9295 8456
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| Fax |
+61-2-9295 8404
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| Organization name |
Garvan Institute
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| Department |
Immunology Program
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| Street address |
384 Victoria St
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| City |
Darlinghurst |
| State/province |
NSW |
| ZIP/Postal code |
2010 |
| Country |
Australia |
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| Platforms (1) |
| GPL1261 |
[Mouse430_2] Affymetrix Mouse Genome 430 2.0 Array |
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| Samples (11)
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| GSM915682 |
NOD B cells, 2h unstimulated, biological replicate 1 |
| GSM915683 |
NOD B cells, 2h unstimulated, biological replicate 2 |
| GSM915684 |
NOD B cells, 2h unstimulated, biological replicate 3 |
| GSM915685 |
NOD B cells, 2h anti-IgM stimulation, biological replicate 1 |
| GSM915686 |
NOD B cells, 2h anti-IgM stimulation, biological replicate 2 |
| GSM915687 |
NR4 B cells, 2h unstimulated, biological replicate 1 |
| GSM915688 |
NR4 B cells, 2h unstimulated, biological replicate 2 |
| GSM915689 |
NR4 B cells, 2h unstimulated, biological replicate 3 |
| GSM915690 |
NR4 B cells, 2h anti-IgM stimulation, biological replicate 1 |
| GSM915691 |
NR4 B cells, 2h anti-IgM stimulation, biological replicate 2 |
| GSM915692 |
NR4 B cells, 2h anti-IgM stimulation, biological replicate 3 |
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| Relations |
| BioProject |
PRJNA159413 |
| Supplementary file |
Size |
Download |
File type/resource |
| GSE37294_RAW.tar |
39.5 Mb |
(http)(custom) |
TAR (of CEL) |
| Processed data included within Sample table |
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