The separated single cells of spleen from WT or WAS-KO mcie, the gel beads with barcoded sequences and the oil droplets were put into the machine together. Thee gel beads precisely captured the target cells, then the target cells were lysed, thus releasing a large quantity of RNA and barcoded sequences for further reverse transcription. Every cell and transcript were barcoded uniquely by using a unique molecular identifier (UMI). Next, libraries were established and sequenced from the cDNA and the 10 × barcodes were adopted to relate individual reads back to the individual partitions. The single cell 3' protocol generating libraries were then sequenced using an Illumina HiSeq4000 or Novaseq sequencer. Transcripts were mapped to the mm10-3.0.0 reference genome.