Single-Cell RNA Sequencing with 10x Genomics

Cell suspensions were loaded on a chromium single-cell controller (10× Genomics, San Francisco, CA, USA) to generate single-cell gel beads in emulsion (GEMs). A Chromium Next GEM Single Cell 3ʹ GEM, Library & Gel Bead Kit v3.1 (1000121; 10× Genomics, Pleasanton, CA, USA) and Single-Cell B Chip Kit (1000127; 10× Genomics) were used according to the manufacturer’s protocol. As previously described, single-cell droplet collection, cDNA amplification, and sequencing library preparation were performed [46 (link),47 (link)]. The cDNA libraries were sequenced on an Illumina Novaseq6000 sequencer with a sequencing depth of at least 30,000 reads per cell and a pair end of 150 bp (PE150; Capitalbio Technology Corporation, Beijing, China).