title: GSM9162479: 8-week old C57BL/6J mouse lung Ly6G⁻Ly6C⁻CX3CR1⁺CD11b⁺SIGLEC-F⁻CD16.2^hi non-classical monocytes rep2 - GEX; Mus musculus; RNA-Seq
library_construction_protocol: Eight-week-old C57BL/6J mice (Mus musculus) were treated intranasally with 10 µg of the cysteine protease papain in a 100 µL volume. After 24 hours, lungs were processed into a single-cell suspension, and two distinct cell populations were isolated by fluorescence-activated cell sorting (FACS): Ly6G⁺Ly6C⁺CX3CR1⁺CD11b^hi SIGLEC-F⁻ macrophages (Ly6G⁺ MΦ) and Ly6G⁻Ly6C⁻CX3CR1⁺CD11b⁺SIGLEC-F⁻CD16.2^hi non-classical monocytes. The sorted cells were subjected to scRNA-seq to characterize their transcriptional profiles. Cell suspension and 10x barcoded gel beads were loaded into the 10x Chromium™ Single Cell Chip G (PN 1000120) to capture single cells in nanoliter-scale oil droplets using the 10x Genomics Chromium X Controller with Chromium Next GEM Single Cell 3' v3.1 reagents (PN-1000268). The single-cell gene expression libraries were constructed following the 10x Genomics 3' Next GEM 3.1 kit instructions. The libraries were then sequenced on an Illumina NovaSeq 6000, targeting 20,000 reads per cell.