title: Mouse_Th0_48h_2
organism: Mus musculus
description: Protocols: T cells from blood (human) and mouse (spleen) CD4+CD62L+ naive T cells were purified from spleens of wild-type C57BL/6JAX adult (6 - 8 weeks) mice using the CD4+CD62L+ T Cell Isolation Kit II (Miltenyii #130-093-227). Cell culture plates were coated with anti-CD3e antibody (1 ug/ml, eBioscences #16-0031-81) in 1X DPBS (Gibco) at 4°C overnight. Purified naive T cells were seeded at a concentration of 1 M cells/ml on the coated plates in IMDM (Gibco) with 10% heat-inactivated FBS (Sigma), supplied with 5 ug/ml anti-CD28 (eBioscience #16-0281-86) with (Th2) or without (Th0) 10 ng/ml mouse recombinant IL-4 (R&D Systems #404-ML-050). Cells were cultured in plates for up to 72 hours. Total RNA was purified by Qiagen RNeasy Mini Kit according to manufacturer's instruction, and concentration was determined by a Nanodrop. A total of 500 ng RNA was used to prepare sequencing libraries using KAPA Stranded mRNA-seq Kit (KAPA #07962193001) according to manufacturer's instructions.
attributes.tag: ENA first public
attributes.value: 2017-12-12
attributes.tag: ENA last update
attributes.value: 2017-12-12
attributes.tag: External Id
attributes.value: SAMEA104423674
attributes.tag: INSDC center alias
attributes.value: Sanger Institute
attributes.tag: INSDC center name
attributes.value: Sanger Institute
attributes.tag: INSDC first public
attributes.value: 2017-12-12T17:01:54Z
attributes.tag: INSDC last update
attributes.value: 2017-12-12T11:42:20Z
attributes.tag: INSDC status
attributes.value: public
attributes.tag: Submitter Id
attributes.value: E-MTAB-6300:Mouse_Th0_48h_2
attributes.tag: age
attributes.value: 6 to 8
attributes.tag: broker name
attributes.value: ArrayExpress
attributes.tag: cell type
attributes.value: T-helper 0 cell
attributes.tag: common name
attributes.value: house mouse
attributes.tag: disease
attributes.value: normal
attributes.tag: genotype
attributes.value: wild type genotype
attributes.tag: individual
attributes.value: 2
attributes.tag: organism part
attributes.value: spleen
attributes.tag: sample name
attributes.value: E-MTAB-6300:Mouse_Th0_48h_2
attributes.tag: stimulus
attributes.value: anti-CD28 (5 microgram per milliliter)
attributes.tag: strain
attributes.value: C57BL/6J