Isolation of Liver Non-Parenchymal Cells

Liver non-parenchymal cells were isolated as previously described^{44 (link),45 (link)}. Briefly, the liver was washed with EGTA solution (Sigma Aldrich, St. Louis, MO) and digested with type 1 collagenase (Worthington Biochemical Corporation, Lakewood, NJ) through the portal vein. Hepatic non-parenchymal cells were separated from hepatocytes by centrifugation. HSCs were separated from non-parenchymal cells using Optiprep gradient solution (Sigma-Aldrich, St. Louis, MO). Monocytes and Kupffer cells were isolated using the FACS Aria III device (BD Bioscience, San Jose, CA) and liver sinusoidal endothelial cells (LSECs) were separated using magnetic-activated cell sorting (MACS) with anti-CD146-PE and anti-PE beads (Miltenyi Biotec, San Diego, CA) following manufacturer’s protocol.