GSM8143826: P1T; Homo sapiens; RNA-Seq
For PSCC tumor and normal adjacent tissues, whole single cells were centrifuged for 5 minutes at 500g and were resuspended in PBS with 0.04% BSA. Cells were then counted using the Countess® II Automated Cell Counter (Thermo Fisher) with final viability of > 85%. For experiments requiring cell sorting, cells first were incubated with viability factor (Zombie NIR™ Fixable Viability kit) in 1×PBS for 15 minutes, followed by washing with MACS buffer (1×PBS with 0.5% FBS and 2 mM EDTA) and centrifugation at 500g for 5 minutes and stained against anti-human CD45 antibody in MACS buffer for 30 minutes. Cells were then centrifuged again, resuspended in MACS buffer, and sorted using a BD FACSAria™ III cell sorter. Sort cells were collected directly in 1×PBS with 0.04% BSA. Chromium Single Cell 3ʹ Reagent Kits v3 (10x Genomics) for cells isolated from tumor and normal adjacent tissues. Chromium Single Cell 5' Library and Gel Bead Kit (10x Genomics) for CD45+ immune cells by FACS sorting.