GSM9018401: non-tumor bearing GFP control; Mus musculus; RNA-Seq
Livers were subjected to a two-step collagenase digestion 2 weeks post-viral infection. Briefly, after anesthetizing the mice, a 27.5” gauge catheter was placed into the inferior vena cava and 0.3 mg/mL of collagenase II (Worthington) was perfused through the liver to promote digestion. Livers were then placed in DMEM/F12 medium (Corning) supplemented with 5% FBS (Biowest). This was then passed through a 100 μm filter followed by a 70 μm filter and centrifuged at low speed (400 rpm for 3 minutes) and the pellet was washed twice. Hepatocyte viability, determined by trypan blue staining, was typically >80%. Isolated hepatocytes and purified non-parenchymal cells were used for sc-RNA-Seq. Cells isolated from 1 individual control and 1 individual Glul-KO mouse were sequenced in separate wells on the flow cell. At minimum, 10,000 cells were sequenced at 20,000 reads per cell. Single cell sequencing was performed on NextSeq 2000 and 5PrimeV3 library prep was conducted according to standard methods. 10X snRNA-seq 5PrimeV3 library preparation