GSM8578772: CD94-KO, scRNA-seq; Mus musculus; RNA-Seq
To obtain CD45+ tumor‑infiltrating lymphocytes (TILs) from WT B6 or CD94-KO mice, the TILs were incubated with an anti-CD45 mAb for 30 min at 4°C. Subsequently, 7AAD was added to the CD45+ cell suspension, which was subsequently sorted using a FACSAria Fusion (BD Biosciences, San Diego, USA). The purity of the sorted cell populations was >  90%. Purified CD45+ live cells were labelled with 0.4% Trypan blue to determine their viability (> 90% before processing for scRNA-seq). Single-cell barcode scRNA-seq libraries were generated using the Chromium Single Cell 3' Library (v2 chemistry; Nugen) according to the manufacturer's standard protocol. The libraries were sequenced on a NovaSeq 6000 (Illumina). Gene-barcode matrices were generated by counting UMIs and filtering non-cell-associated barcodes.