scRNA-seq of mus musculus: adult male mesentary lymph node
GPR55KO mice with/without O-1602 treatment, 0.4 mg/kg i.p. for 3 hours), mLN with fat tissues carefully removed were collected and pooled from two independent mice. Single-cell suspension was obtained by High Activity Mouse Intestine Tissue Enzymatic Digestion Kit (RWD, DHIE-5007) in accordance with the manufacturers protocol. DCs were enriched using a two-step magnetic selection procedure. In brief, CD3+, CD19+, and NK1.1+ cells in single-cell suspension were partially removed using magnetic beads of STEMCELL EasySep Mouse CD25 Regulatory T Cell Positive Selection Kit (#18782). CD45+ cells containing total DCs were isolated from the above remains by the same STEMCELL magnetic beads (#18782), followed by being labelled with Anti-Mouse MHC-H2 Class I barcode sample tags (BD Mouse Single-Cell Multiplexing Kit, 626545). Single cells from various groups were mixed at a 1:1 ratio. Dead cells were excluded by DAPI (Beyotime, C1006) staining and high activity cells were sorted by FACS for single-cell capture and library preparation.