GSM5769214
Splenocytes were loaded on a Chromium Controller (10x Genomics, Pleasanton) at a concentration of 10^3 cells/µL per sample, to generate Gel Bead-In EMulsions (GEMs). Each GEMs is wrapped with a beads and a cell, and then cell lysis and reverse transcription reaction are carried out in GEMs to synthesize cDNA with barcode sequence. PCR amplification is carried out with cDNA. After amplification, quality inspection is carried out for the amplified products. Barcoded sequencing libraries were constructed using the Chromium Single Cell 3' Reagent Kits v3 (10x Genomics) according to the manufacturer's instructions. Following library preparation, sequencing was performed with paired-end sequencing of 150 nt at each end using one lane of the NovaSeq 6000 per sample. Raw sequencing data were processed using Cell Ranger (10x Genomics; v6.0.1). The CellRanger function “count” was used to align raw reads with the STAR aligner against the mm10 reference genome. After counting nonredundant unique molecular identifiers (UMIs), single-cell digital-expression matrices were obtained. sc-RNASeq