GSM8986565: CD45+ WT GEX; Mus musculus; RNA-Seq
Lungs were minced and digested, and flow cytometric sorting (BD FACSAria II) was used to prepare circulating CD45.2+ leukocytes from WT and Plaur–/– mice. Approximately 16,000 live cells at or above 4×10^5 live cells/mL with 70% or higher viability were loaded into the Single Cell Chip followed by forming 10,000 single cell emulsion in Chromium Controller (10x Genomics). The first strand of cDNA for antibody derived transcripts (ADT) and gene derived transcripts is synthesized according to manufacturer's instructions (10x Genomics, catalogue #PN-1000268). Then the cDNA is amplified according to the Total A-seq instruction (BioLegend) for samples with Total-seq A antibodies and the Chromium single cell 3′ library and gel bead kit v3.1 (Protocol CG000315, 10x Genomics). The Gene Expression and the ADT Libraries were prepared separately following manufacturer's instructions. The two types of libraries were mixed at 10:1 molar ratio (Gene Expression library to ADT library) and sequenced by the NIEHS Epigenomics and DNA Sequencing Core Laboratory on NovaSeq 6000 (Illumina) with paired-end sequencing (Read1: 28;  Read2: 90). A total of 288,184,128 reads were obtained.