title: Single-cell RNA sequencing data of Ly49⁺ CD8 T cells treated with anti-CD80/86 antibody and control Ig
design: Single-cell RNA sequencing data of Ly49⁺ CD8 T cells treated with anti-CD80/86 antibody and control Ig
library_name: S2P1N26_p
library_construction_protocol: B6 splenocytes were obtained from naïve mice or after MLR in the presence or absence of anti-CD80/86 mAb. Anergic B6 splenocytes obtained after MLR with anti-CD80/86 mAb were added into MLR of Ly49+ cell-depleted B6 splenocytes with irradiated LPS-stimulated BALB/c splenocytes, then live cells were prepared on day 3 and stained with FITC-conjugated anti-mouse CD3 mAb (17A2), APC/Cy7-conjugated anti-mouse CD8α mAb (53-6.7), and PE-conjugated anti-mouse Ly49 mAb (14B11).Single Ly49+ CD8+ T cell was dropped into every well on 96 well plates preloaded with lysis buffer containing NP-40 and RNase inhibitor using FACSmelody (BD Biosciences). After a single freeze and thawing cycle, cDNAs were synthesized from whole cell lysate and amplified by one-step RT-TS-PCR method (Repertoire Genesis, Osaka, Japan) using PrimeScriptⅡ High Fidelity One-Step RT-PCR Kit (Takara Bio, Kusatsu, Japan), adapter-added oligo dT primers, adaptor-added template-switch oligos and adaptor primers.  Sequencing libraries were generated using KAPA HyperPlus Kit (Roche Diagnostics, Basel, Switzerland) and NEBNext Multiplex Oligos for Illumina (New England Biolabs) following the manufacturer's recommendations. 
identifiers.namespace: European Bioinformatics Institute
identifiers.id: E-MTAB-16148:S2P1N26_p