title: GSM9209898: BRI-1986_HumanLN_GEX_HTO_Library; Homo sapiens; RNA-Seq
library_construction_protocol: (BRI-2314, BRI-2316, BRI-2322, BRI-2324) Live CD4+CD19-CXCR5+ cells were sorted from spleens of the NP-OVA immunized mice based on GFP and TdTomato expression into Tfr (GFP-TdTomato+) and ExTfr (GFP-TdTomato+). Cells were stained with TotalSeq-C Hashtags (Biolegend) and pooled in a single sample before loading. (BRI-2146, BRI-2148) Live CD4+CD19-CXCR5+GITR+ Tfr cells were sorted from the inguinal lymph nodes of the NP-OVA immunized mice. Cells were stained with TotalSeq-C Hashtags (Biolegend) and pooled in a single sample before loading. (BRI-1986) Live CD4+CD19-CXCR5+ cells were isolated by FACS-sort from cryopreserved human peripheral blood mononuclear cells (n=3) and iliac lymph node mononuclear cells  (n=3) obtained from six different donors. Cells were resuspended in 0.4% BSA in PBS at a concentration of 2,000 cells per μl, then loaded onto a single lane (Chromium chip K, 10X Genomics) followed by encapsulation in a lipid droplet (Single Cell 5′kit V2, 10X Genomics) followed by cDNA and library generation according to the manufacturer's protocol. 5' mRNA library was sequenced to an average of 50,000 reads per cell, V(D)J library and HTO (Cell Hashing antibodies) library sequenced to an average of 5,000 reads per cell, all using Illumina Novaseq.