GSM3073365: ILC1_D6_S6; Homo sapiens; RNA-Seq
The three ILC subsets were isolated using flow cytometry. In brief, CD3, CD14 and CD19 cells were MACS (Miltenyi Biotec, Bergisch Gladbach, Germany) depleted and then sorted using a FACS Aria flow cytometer (BD Bioscience, Allschwil, Switzerland). Staining procedure for sorting should be also included here. In brief, lymphocytes were initially gated based on FSC and SSC, and then dead cells were excluded using a Fixable Viability Dye (eBioscience, Frankfurt, Germany). We then gated on the linage-CD127+ viable lymphocytes population, of which the CD161mid/+ cells were the main ILC containing population. The three ILC subsets were finally distinguished based on their differential expression of c-kit, CRTH2 and NKp44. The sorted ILC populations (from N = 6 individuals) were immediately re-sorted (2-round sorting) into 96-w plate, 1000 cells per well or as close as possible. RNA libraries were prepared for sequencing using Clontech SMARTer - Low Input Mammalian kit.
GEO Accession
GSM3073365