|
| Status |
Public on Feb 14, 2018 |
| Title |
Cre^(ER)/Rptor^(fl/fl) pro-B cells untreated replicate 3 |
| Sample type |
RNA |
| |
|
| Source name |
Untreated pro-B cells from Cre^(ER)/Rptor^(fl/fl) mouse
|
| Organism |
Mus musculus |
| Characteristics |
cell population: B220+CD43+IgM- pro-B
strain: C57BL/6 Cre^(ER)/Rptor^(fl/fl)
|
| Treatment protocol |
Pro-B cells from tamoxifen-treated Cre^(ER)/Rptor^(fl/+) and Cre^(ER)/Rptor^(fl/fl) mice were cultured in the presence or absence of 25 ng/ml IL-7 for 24h.
|
| Growth protocol |
Lymphocytes were isolated from bone marrow of mice, and pro-B cells (B220+CD43+IgM-) were sorted on a MoFlow (Beckman-Coulter) or Reflection (i-Cyt).
|
| Extracted molecule |
total RNA |
| Extraction protocol |
RNA was extracted using Qiagen's RNAeasy kit, with on-column DNase digestion included.
|
| Label |
biotin
|
| Label protocol |
One to five nanograms of total RNA were converted into biotinylated cDNA using the NuGEN WTA Pico v2 protocol and the NuGEN Encore Biotin module
|
| |
|
| Hybridization protocol |
Six micrograms of biotinylated cDNA was fragmented and then hybridized for 16 hr at 45C on an Affymetrix Mouse Gene 2.0 ST GeneChip. Following hybridization, GeneChips were washed and stained in the Affymetrix Fluidics Station 450.
|
| Scan protocol |
GeneChips were scanned using the AffymetrixGeneChip Scanner 3000 7G.
|
| Data processing |
Probe signals were quantile normalized and summarized by the RMA algorithm using Partek Genomics Suite software v6.6
|
| |
|
| Submission date |
May 24, 2016 |
| Last update date |
Feb 14, 2018 |
| Contact name |
Geoffrey Neale |
| E-mail(s) |
geoffrey.neale@stjude.org
|
| Organization name |
St Jude Childrens Research Hospital
|
| Department |
Hartwell Center
|
| Street address |
262 Danny Thomas Place
|
| City |
Memphis |
| State/province |
TN |
| ZIP/Postal code |
38105 |
| Country |
USA |
| |
|
| Platform ID |
GPL16570 |
| Series (1) |
| GSE81844 |
B cell development requires discrete regulation by PTEN-PI3K and mTOR pathways and the intricate interplay with IL-7 r-Stat5 signaling |
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