title: iTreg rep2
organism: Mus musculus
description: Protocols: CD4+, CD62L+ cells were purified from DEREG mice with CD4+CD62L+ T Cell Isolation Kit II (Miltenyi Biotec). Cells were seeded into anti-CD3 (1 ug/ml, clone 145-2C11, eBioscience) and anti-CD28 (5 ug/ml, clone 37.51, eBioscience) coated 96-well plates, at a density of 250,000-500,000 cells/ml and cultured in a total volume of 200 ul in the presence of recombinant human TGF-beta1 (20ng/ml, Sigma). The cells were gently removed from the activation plate on day 4. The iTreg culture was subjected to FACS sorting for GFP-positive, propidium iodide-negative cells. Poly-(A)+ RNA was purified from ~500,000 cells using the Oligotex kit (Qiagen). The manufacturers protocol was modified slightly to include additional final elution steps resulting in a larger volume. RNA was purified, reverse transcribed and prepared for sequencing described in http://msb.embopress.org/content/7/1/497
attributes.tag: ENA-FIRST-PUBLIC
attributes.value: 2015-04-05T17:04:12Z
attributes.tag: ENA-LAST-UPDATE
attributes.value: 2018-03-08T17:48:50Z
attributes.tag: External Id
attributes.value: SAMEA2536554
attributes.tag: INSDC center name
attributes.value: EBI
attributes.tag: INSDC first public
attributes.value: 2015-04-05T17:04:12Z
attributes.tag: INSDC last update
attributes.value: 2018-03-08T17:48:50Z
attributes.tag: INSDC status
attributes.value: public
attributes.tag: Submitter Id
attributes.value: E-MTAB-2582:iTreg rep2
attributes.tag: broker name
attributes.value: ArrayExpress
attributes.tag: cell type
attributes.value: induced T-regulatory cell
attributes.tag: common name
attributes.value: house mouse
attributes.tag: sample name
attributes.value: E-MTAB-2582:iTreg rep2
attributes.tag: scientific_name
attributes.value: Mus musculus