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Sample GSM154730 Query DataSets for GSM154730
Status Public on May 18, 2007
Title GITR+ CD4+ cells from C57BL/6 mice transduced with empty MigR1 vector
Sample type RNA
Source name pooled lymph node and spleen
Organism Mus musculus
Characteristics Strain: C57BL/6
Gender: Female Age: 6-8 weeks
Treatment protocol CD25+CD4+ from B6 mice or CD25–CD4+ cells from B6.SJL mice (Ly5.1+), isolated by magnetic bead sorting, were stimulated with 100 ng/ml anti-CD3 and 100 ng/ml anti-CD28 in goat-anti-hamster IgG (Jackson ImmunoResearch) coated 12-well plates. IL-2 (100 U/ml) was added to CD25+CD4+ cells on day 0 and to CD25–CD4+ cells on day 1 of culture. On days 1 and 2 following stimulation, cells were “spinfected” (90 min, 2500 rpm, 37°C) with retroviral supernatants, supplemented with 5 ug/ml polybrene, 10 mM HEPES and IL-2, from transfected phoenix-E cells transfected with MigR1, MigR1-PDE3B or MigR1-PDE3B-H801A. On day 5 of culture, aliquots of cells were stained for Foxp3. Cultures of CD25+CD4+ cells routinely contained >70% Foxp3+ cells, >55% of which were transduced (GFP+), while cultures of CD25–CD4+ cells contained >35% transduced cells. CD25–CD4+ cells were spinfected only on day 1 because retroviral transduction of these cells was more efficient than for TR. Cells transduced with the same vectors were mixed and transferred into TCRalpha-deficient recipients. Three weeks after transfer cells were analyzed by flow cytometry and isolated by FACS sorting for gene expression analysis.
Extracted molecule total RNA
Extraction protocol TelTest Stat-60 RNA extraction.
Label biotin
Label protocol RNA was converted to biotinylated cRNA (2 rounds of amplification) using commercially available reagent kits (MessageAmp II, Ambion, Austin, TX) and were hybridized to Affymetrix mouse 430 2.0 microarrays.
Hybridization protocol Standard hybridization to Affymetrix 430_2.0 arrays.
Scan protocol Standard scanning of Affymetrix 430_2.0 arrays.
Description GITR expression and Foxp3 expression correlated well in these experiments. Transduced cells were dectected by the MigR1 marker gene, GFP. GFP+ GITR+ CD4+ Ly5.1- cells were sorted from recipient mice for RNA.
Data processing GCMRA, Bioconductor
Submission date Jan 10, 2007
Last update date Aug 28, 2018
Contact name Alexander Rudensky
Organization name University of Washington
Department Immunology
Street address 1959 Pacific St
City Seattle
State/province WA
ZIP/Postal code 98195
Country USA
Platform ID GPL1261
Series (2)
GSE7280 Gene expression in peripheral cells: effects of Foxp3 and PDE3B
GSE7773 Foxp3-dependent programme of regulatory T-cell differentiation
Reanalyzed by GSE119085

Data table header descriptions
VALUE TR cells - vector

Data table
1415670_at 9.24704627
1415671_at 8.270998069
1415672_at 10.95554288
1415673_at 7.344598015
1415674_a_at 8.150315084
1415675_at 8.35299233
1415676_a_at 10.54804284
1415677_at 6.692342384
1415678_at 9.177035587
1415679_at 11.70205132
1415680_at 8.075284185
1415681_at 9.118003359
1415682_at 6.543980408
1415683_at 10.49262992
1415684_at 7.2330665
1415685_at 7.994417604
1415686_at 7.053163643
1415687_a_at 9.242509402
1415688_at 9.546719828
1415689_s_at 8.894482798

Total number of rows: 45101

Table truncated, full table size 1026 Kbytes.

Supplementary file Size Download File type/resource
GSM154730.CEL.gz 3.1 Mb (ftp)(http) CEL
Raw data provided as supplementary file
Processed data included within Sample table

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