GSM4932903: J10624_RNA; Homo sapiens; RNA-Seq
CD154+ cells were isolated by MACS and further purified by FACS sorting on a MACSQuant Tyto (Miltenyi Biotec) based on dual expression of CD154 and CD69. Sorted CD154+ T cells were removed from the sorting chamber into pre-coated low-bind collection tubes, 1ml RPMI-1640 medium supplemented with 5% AB Serum was added, and cells were centrifuged for 5 min at 400 x g, 4°C. The supernatant was carefully removed leaving 10-30μl to reach a maximum concentration of 1000 cells /μl. Single-cell suspensions were loaded on a ChromiumChip G (10x Genomics) according to the manufacturer's instructions for processing with the Chromium Next GEM Single Cell 5′ Library and  Gel  Bead  Kit  v1.1. TCR  single-cell  libraries  were subsequently prepared from the same cells with the Chromium Single Cell V(D)J Enrichment Kit, Human T Cell. Chromium Next GEM Single Cell 5′ Library