characteristics: ERC000011
characteristics: ENA-CHECKLIST
characteristics: 2022-10-01T16:26:42Z
characteristics: ENA-FIRST-PUBLIC
characteristics: 2022-10-01T16:26:42Z
characteristics: ENA-LAST-UPDATE
characteristics: SAMEA111379123
characteristics: External Id
characteristics: Department of Dermatology, Inselspital, Bern University Hospital, University of Bern, Bern, Switzerland
characteristics: INSDC center name
characteristics: 2022-10-01T16:26:42Z
characteristics: INSDC first public
characteristics: 2022-10-01T16:26:42Z
characteristics: INSDC last update
characteristics: public
characteristics: INSDC status
characteristics: ERS13476106
characteristics: SRA accession
characteristics: E-MTAB-12197:2_12h_D
characteristics: Submitter Id
characteristics: 40
characteristics: year
characteristics: age
characteristics: ArrayExpress
characteristics: broker name
characteristics: T helper 9 cell
characteristics: cell_type
characteristics: human
characteristics: common name
characteristics: Protocols: Peripheral Blood Mononuclear Cells (PBMC) were isolated by Ficoll-Plaque Plus (GE Healthcare, UK) density gradient centrifugation. Human CD4+ T cells were isolated from PBMC by EasySep positive selection kit (Stemcell Technologies) according to manufacturer’s instruction. Positively selected CD4+ T cells were washed with PBS and stained for subsequent Th cell subset sorting. Effector memory Th9 cells (CXCR3-CCR8+CCR6-CCR4+) were sorted to over 90% purity according to their expression of chemokine receptors from CD45RA-CD25-CD8-CD3+ cells. Single cell Th9 clones were directly sorted into 96well plate according to their expression of chemokine receptors. Culture medium consisted of PRMI-1640 with Hepes (GIBCO) supplemented with 5% of heat-inactivated human serum (Swiss Red Cross, Basel, Switzerland), 2mM L-Glutamine (Biochrom), 50U/ml penicillin and 50μg/ml streptomycin (Bioconcept) and 50 IU/ml IL-2 (Roche). T cells were cultured at a density of 0.25-1x105cells/96 well plate in a total volume of 200μL cell culture medium Th9 cell clones were incubated with GW9662 for 48 hours and activated with αCD3/2/28 for 12 hours Total RNA was isolated with RNAeasy kit (Qiagen) Illumina TruSeq Stranded mRNA protocol (Directional, first strand)
characteristics: description
characteristics: adult
characteristics: developmental stage
characteristics: normal
characteristics: disease
characteristics: Homo sapiens
characteristics: organism
characteristics: Homo sapiens
characteristics: organism
characteristics: blood
characteristics: organism part
characteristics: 2
characteristics: replicate
characteristics: Homo sapiens
characteristics: scientific_name
characteristics: male
characteristics: sex
characteristics: antibodies against CD3, CD2, and CD28 (12 hours)
characteristics: stimulus
characteristics: 2_12h_D
characteristics: title