GSM5725953: T cells HTO; Mus musculus; RNA-Seq
Cells were extracted from pooled spleens isolated from seven 2-week-old- and two 5-week-old-WT mice, nine 1-week-old-, eleven 1.5-week-old-, six 2-week-old-, five 2.5-week-old-, three 3-week-old-, and one 5-week-old-LatY136F mice, and T cells were enriched using the EasySep Mouse T Cells Isolation Kit (Stemcell). Each purified T cell sample was then labeled with a specific Hashtag (TotalSeq™ anti-mouse Hashtag antibody #A0301-A0308, Biolegend), a mix of 6 calibrated TotalSeq™-A anti-mouse specific antibodies (CD27, CD25, CD39, TCRb, PD-1, CD103), and a cocktail of 6 fluorescently-labeled anti-mouse antibodies plus a viability dye. Cells were then sorted as live singlets, CD11b– IA/IE–CD90.2+ CD6+ expressing either CD4+ or CD8+. After sorting, cells were washed in PBS - 0.04% BSA as recommended by the 10xGenomics sample preparation protocol and kept on ice until counting was performed. 2 to 3 samples were mixed at a concentration of 1200 cells /µl and loaded in 2 to 3 lanes at a maximum concentration of 12,000 cells per lane and run on a Chromium-controller (10X Genomics, Pleasanton). Single-cell RNA-Seq libraries, HTO libraries and ADT libraries were generated using the Chromium Single Cell 3′ V3 kit (10X Genomics) according to the manufacturer's instructions.