GSM5718097: F3_IL5CreΔIfnγr1  IL33 + IFNγ; Mus musculus; RNA-Seq
Approximately 10,000-20,000 ILC2s from lungs and adipose tissue were sorted int0 100-200ul of lysis/binding buffer (Qiagen) using a BD FACSAria II sorting system with a 70um nozzle size and 4 way-purity sort mode. ILC2 were sorted from IL5Cre-Tdtomato Red5 mice as live, lineage negative, CD45+,Thy1.2+, Tdtomato+. Total RNA was isolated using QIAGEN RNeasy Plus Micro kit Gene expression profiling was carried out using a 3' Tag-RNA-Seq protocol. Barcoded sequencing libraries were prepared using the QuantSeq FWD kit (Lexogen, Vienna, Austria) for multiplexed sequencing according to the recommendations of the manufacturer (Lexogen). The fragment size distribution of the libraries was verified via micro-capillary gel electrophoresis on a Bioanalyzer 2100 (Agilent, Santa Clara, CA). The libraries were quantified by fluorometry on a Qubit fluorometer (LifeTechnologies, Carlsbad, CA), and pooled in equimolar ratios. Up to forty-eight libraries per lane were sequenced on a HiSeq 4000 sequencer (Illumina, San Diego, CA).
GEO Accession
GSM5718097