GSM9212053: T_cells_plus_PD1; Mus musculus; RNA-Seq
The day of library preparation, samples were thawed and like samples were combined. All cells were stained with viability dye, CD4, CD8 and Thy1.1 antibodies for sorting. Samples were acquired with an Aria2-2 or a Symphony sorter (BD). Live engineered T cells (live/dead- CD8+ Thy1.1+), endogenous CD8 T cells (live/dead- CD8+ Thy1.1-), endogenous CD4 T cells (live/dead- CD4+ Thy1.1-), and the remaining live cells (live/dead-) were sorted and each cell population was stained with a unique cell hashing antibody (Hash#1-4). Samples were pooled after hashing and stained with the TotalSeq-C Custom Mouse Panel (Biolegend, Cat:900002538). A gene expression library was generated (10x Genomics Next GEM Chromium Single Cell 5' Gene Expression v2 and Mouse TCR amplification Kits) according to Manufacturer's protocol