GSM5328180: FoxP3 GFP+ lymph node - uninjured - GEM; Mus musculus; RNA-Seq
viable lymph node FoxP3 GFP+ cells were sorted from injured or uninjured Foxp3DTR mice at 7 days after injury. The sorted FoxP3 GFP+ cells were washed and resuspended in sorting buffer at a cell concentration of 1000 cells/µL. About 17,000 mouse cells were loaded onto a 10x Genomics ChromiumTM instrument (10x Genomics) according to the manufacturer's recommendations. The scRNAseq libraries were processed using ChromiumTM single cell 5' library & gel bead kit (10x Genomics). Matched scTCRseq libraries were prepared using 2 µL of post cDNA amplification material and Chromium Single Cell V(D)J Enrichment Kit, Mouse T Cell. Quality controls for amplified cDNA libraries and final sequencing libraries were performed using Bioanalyzer High Sensitivity DNA Kit (Agilent). The sequencing libraries for scRNAseq and scTCRseq were normalized to 4nM concentration and pooled using a volume ratio of 4:1. The pooled sequencing libraries were sequenced on Illumina NovaSeq S4 300 cycle platform. The sequencing parameters were: Read 1 of 150bp, Read 2 of 150bp and Index 1 of 8bp.