GSM6564268: Pool of myeloid cells from ILC2-proficient mice and epithelial cells from ILC2-deficient mice; Mus musculus; RNA-Seq
Lung tissue was digested using Liberase TM and DNaseI to obtain single-cell suspensions. Cells were subsequently incubated with fluorochrome-conjugated antibodies and epithelial cells were sorted by FACS as live CD45lo/- EpCAM+ cells, myeloid cells were sorted as live CD45+ (CD11b and/or CD11c)+. Epithelial cells from ILC2-proficient mice were pooled with myeloid cells from ILC2-deficient mice and vice versa. These cells were then applied to the 10X Genomics platform using the Chromium™ Single Cell 5' Library & Gel Bead Kit v2 (10x Genomics) following the manufacturer's instructions. The purified cDNA was amplified using 12 cycles of PCR and then used for 5' gene expression (GEX). Through fragmentation, adapter ligation and index PCR the final libraries were obtained. The quality of single cell 5' GEX was assessed by Qubit quantification and Bioanalyzer fragment analysis (HS DNA Kit, Agilent).The sequencing was performed on a NextSeq2000 device (Illumina) with the recommended sequencing conditions for 5' GEX libraries (read1: 26nt, read2: 90nt, index1: 10nt, index2:10).