GSM8385068: MCD+EMPA, replicate 2, scRNAseq; Mus musculus; RNA-Seq
Mouse liver tissue was digested, passed through a 100 mm cell strainer, and subjected to red blood cell lysis. Cells were incubated with CD45 antibody at 4°C for 30 minutes, followed by incubation with DAPI (Thermo Scientific, 62248, 1:1000) for 5 minutes. Cells were resuspended in 1640 medium and filtered through a 40 mm cell strainer. Pre-prepared collection tubes containing 1640 medium with 10% FBS were sent to the sorting room. The sample tubes were loaded into the sample chamber for target cell sorting. Sorted cells were mixed, and a portion was reanalyzed to record flow cytometry data. Cells were quality controlled to ensure >80% viability, washed, and resuspended to 700-1200 cells/µL. Libraries were constructed for 3' gene expression by fragmentation, end repair, adapter ligation, and indexing. Sequencing was performed on the Illumina Novaseq 6000 platform with PE150 mode, achieving a depth of 50,000 read pairs per cell.