GSM9757415: Single-cell RNA-seq of mouse Siglec-G BKO Tfh cells; Mus musculus; RNA-Seq
Single-cell suspensions were prepared by mechanically dissociating harvested organs. Cells were stained with fluorophore-conjugated antibodies against Tfh/Tfr cell surface markers. Viable Follicular T helper (Tfh) and T follicular regulatory (Tfr) cells were isolated via fluorescence-activated cell sorting (FACS) on a high-speed cell sorter. Single cells were partitioned and captured into nanoliter-scale Gel Beads-in-emulsion (GEMs) using the 10x Genomics Chromium Controller. Single-cell RNA-seq libraries were constructed using the Chromium Single Cell 3' Reagent Kits (v3/v3.3) according to the manufacturer's user guide. Briefly, GEMs were broken, cDNA was amplified, fragmented, end-repaired, A-tailed, and ligated with sequencing adapters before final library amplification.