GSM8820885: WT2CCL4; Mus musculus; RNA-Seq
Livers were perfused with HBSS and then EMEM-containing collagenase type 4, 40-mg pronase, and 25 mg/mL of DNAase 1. Suspension was centrifuged to isolate hepatocytes in pellet and the supernatant was saved for non-parenchymal cells. The suspension was subjected to purification with 20% percoll gradient. The purified hepatocytes were added to final non parenchymal cells suspension, and the final cell suspension (1200/μL and 100 μL in total) was prepared in phosphate buffered saline containing 0.04% bovine serum albumin and were submitted to Yale Center for Genome Analysis. Cells were subjected to 10X chromium using 3′ gene expression V3kit for library preparation according to manufacturer's guidelines and were sequenced using Novaseq. The count matrix was generated using 10X Genomics Cell Ranger software. The barcode matrices from Cell Ranger output were demultiplexed using Seruat pipeline for QC. The liver sections from rest of the mice were subjected to Sirius red staining to assess the grade of fibrosis. Cells were subjected to 10X chromium using 3′ gene expression V3kit for library preparation according to manufacturer's guidelines and were sequenced using Novaseq. The count matrix was generated using 10X Genomics Cell Ranger software. The barcode matrices from Cell Ranger output were demultiplexed using Seruat pipeline for QC.