title: GSM6737816: Macrophages, ILC2, Mock mice, replicate 2; Mus musculus; RNA-Seq
library_construction_protocol: From ex vivo co-culture, cells were harvested using Cell Dissociation Buffer (ThermoFisher) for 15 min in 37°C and then macrophages (FSC-Ahigh, CD11c+ living cells) were sorted using FACS and kept in TRIzol at -80°C. RNA from macrophages was extracted using the RNeasy Mini kit (Qiagen) according to the manufacturer's instructions, and quality was assessed on Agilent RNA 6000 Pico. cDNA was prepared using SmartSeq HT (1ng). RNA-Seq libraries were prepared from 150ng (5μl at 30pg/μl), using the Illumina Nextera XT DNA Library Preparation Kit. Libraries were quantified and normalized by qPCR. Libraries were finally sequenced using Illumina NovaSeq6000 and bioinformatics analysis was performed. Approximately 25 × 106 75-base single-end reads were generated per sample.