Single cell RNA sequencing
These single cells were loaded on a Chromium Controller instrument to generate single cell gel bead-in emulsions. As the single cells lysed, the gel beads automatically dissolved and released a large number of barcode sequences, followed by reverse transcription of the mRNA to generate cDNA with barcode and UMI information8,9. The sequencing library was constructed after quality inspection and amplification of cDNA, and then sequenced on an Illumina NovaSeq 6000 at a depth of 400 M reads. Using Cell ranger software (10X Genomics, version 2.1.1), the data of raw basecall files were split, counted, combined mathematically, and reanalyzed. Cells with less than 10% UMI in whole genes were filtered out.