title: GSM3291234: WildType CD4-Rep2; Mus musculus; RNA-Seq
library_construction_protocol: WT, Gli2ΔN2 and Gli2ΔC2 mice (6 from each strain) were sensitised on the shaved abdomen with 3% of the hapten 4-ethoxymethylene-2-phenyloxazolin-5-one (Oxazolone, Sigma), and then challenged topically on the ears with five doses of 0.6% Oxazolone on days 5, 7, 9, 11 and 13 and sacrificed and analysed on day 14. To release leukocytes from the whole ear tissue, ears were split into dorsal and ventral halves and digested for 30min at 37°C with 1mg/ml Collagenase D (Roche) and 0.5mg/ml DNase I (Roche) in RPMI (Life Technologies) containing 2% FCS (Life Technologies) and 2% Penicillin/Streptomycin (Life Technologies). Tissues were filtered through a 70μm cell strainer (VWR) and washed in RPMI containing 10% FCS and 5% Pen/Strep, spun at 1400rpm for 10min and counted with haemocytometer (Hawksley). Skin cell suspensions from two groups of three mice each from WT, Gli2ΔN2 and Gli2ΔC2 mice were obtained. Cells were stained and live (ZA-) CD4 T cells (CD45+CD3+γδTCR-CD4+CD8-) and CD8 T cells (CD45+CD3+γδTCR-CD4-CD8+) were sorted into Trizol LS reagent (Thermo Fisher Scientific). RNA was extracted from these cells. RNA libraries were prepared for sequencing using standard Illumina protocols
attributes.tag: GEO Accession
attributes.value: GSM3291234