GSM7229996
Liver cells were isolated from both wild-type (WT) and N-LKO mice fed WD obtained from the Yale Liver Facility Center using a multi-step mechanism. The mice were anesthetized with Isoflurane and attached to a Styrofoam tray, and a ligature was placed around the mid-portal vein and IVC, after which the abdomen was wet with 70% ethanol. The portal vein was cannulated with a 22 G catheter, and perfused with Hanks A followed by Hanks B with collagenase. The liver was removed, and the hepatocytes were released by shaking. The resulting cell suspension was filtered through a 40 μm mesh, and the cells were pelleted by centrifugation. The cell pellet was then resuspended in L-15 media. Samples were prepared according to the instruction of 10x Genomics Single Cell 3, Reagent Kits v3. Single cell suspensions in RT Master Mix were loaded onto the 10x Genomics Single Cell A Chip to convert poly-adenylated mRNA into barcoded cDNA. Barcoded cDNA was amplified by PCR to generate a sufficient mass for library construction.