GSM4630629: NKT_Ja18_b8; Mus musculus; RNA-Seq
Tissue preparation and cell isolation were performed as previously described. Briefly, the livers were perfused with PBS before extraction to remove the blood cell. Then, the livers were minced and filtered through a cell strainer (40 μm; BD Biosciences). Leukocytes were isolated from the liver cell suspensions by density fractionation using discontinuous 40-70% (vol/vol) Percoll (GE Healthcare) gradients, followed by red blood cell lysis before filtration. Then NKT cells were enriched by cell sorting (FACS) of NK1.1+CD3e+ cells and subjected to scRNA-seq. First, individual cells were manually picked, lysed and subjected to first-strand cDNA synthesis as reported. Then, second-strand cDNA was synthesized, amplified and fragmented. RNA-seq libraries were prepared according to the instruction manual of the KAPA Hyper Prep Kit (KAPA Biosystems). Finally, the libraries were checked, pooled together and sequenced with paired-end 150-bp reads on an Illumina HiSeq X-Ten platform.
GEO Accession
GSM4630629