title: GSM7211461: Cartridge 4; Homo sapiens; RNA-Seq
library_construction_protocol: Fresh whole blood was collected from patients and HDs. PBMCs were isolated by density gradient centrifugation. Leukocytes were labeled with Aqua LIVE/DEAD (Thermo Fisher Scientific) stain to remove non-viable cells, and with the following antibodies against: CD45RA-APC-H7, CD3-BV650 and CD4-R718 (BD Biosciences) antibodies. CD45RA-CD3+CD4+ TLs were isolated from PBMCs by flow cytometry sorting. Targeted scRNA-seq analysis was performed with the BD Rhapsody Single-Cell Analysis System (BD Biosciences), according to the manufacturer's instructions. Each sample was stained with 20 BD AbSeq antibodies (Supplemental Table 5). The BD Human Single-Cell Multiplexing Kit was used to multiplex up to seven subjects per Rhapsody cartridge. For library construction, the samples were pooled before cartridge loading. The BD Rhapsody Immune Response Targeted Panel for Humans was used to assess mRNA levels for 399 genes (#633750).