GSM7885253: B; Homo sapiens; RNA-Seq
Ex ILC2s, ILCs from the blood of healthy donors (HD ILCs), ILCs from the blood of patients with AML (AML ILCs), and human CD45+ cells from the bone marrow of NSG mice (Traced ILC2s) were sorted using BD FACSAria Cell samples containing 3,000-6,000 cells were captured using a 10 × Genomics Chromium controller and the 10 × V3.1 Single Cell 3' Solution kit (Chromium Next GEM Single Cell 3' Regent kits V3.1, Cat# PN-1000268). All protocols were followed by the manufacturer's instructions. The final sequencing libraries were assessed for library size using a High Sensitivity DNA Chip (Agilent, Cat# 5067-4626). Library concentration was determined using a Qubit High Sensitivity DNA assay Kit (ThermoFisher Scientific, Cat# Q32854). Sequencing was conducted using the Illumina NovaSeq 6000 platform with the paired-end setting, involving 28 cycles of read1, 101 cycles of R2, 10 cycles of index i7, and 10 cycles of index i5 read. The sequencing kit used was S4 Reagent kit v1.5 (Illumina, Cat# 20028313) at Tgen. Demultiplexing, genome alignment, and feature-barcode matrix generation were performed using the 10 × Genetics Cell Ranger software pipeline. 10x genomics Single Cell RNA-seq