Anti mouse fcγrii 3

To purify KCs, Ly6C^hi and Ly6C^low IMs, liver NPCs were incubated with normal rat serum (Sigma) and anti-mouse FcγRII/III (Becton Dickinson, Franklin Lakes, NJ, USA) to minimize nonspecific antibody binding. Subsequently, the cells were stained with anti-CD45, anti-Ly6C, anti-Ly6G, anti-CD19, anti-SiglecF (Becton Dickinson) and anti-F4/80, anti-CD11b, anti-NK1.1 and anti-CD3 (eBioscience, San Diego, CA, USA), and sorted using a BD FACSAria II Cell Sorter (BD Bioscience, San Jose, CA, USA).

Isolated liver NPCs were incubated with1 μl of anti-mouse FcγRII/III (Becton Dickinson, Franklin Lakes, NJ) to minimize non-specific antibody binding. The cells were then stained with anti-mouse CD45-V655 (eBioscience, 15520837), F4/80-APC/Cy7 (Biolegend, 123118), Ly6C-APC (BD Pharmingen, 560595), Ly6G-V450 (BD Pharmingen, 560603), CD146-PerCP-Cy5.5 (BD Pharmingen, 562134), CD44-PE (BD Pharmingen, 553134), anti-His-FITC (abcam, ab1206). In some experiments, cells were incubated with 2 μg rmChi3l1 for 2 hr before antibody staining. The cells were analyzed on a CytoFLEX LX Flow Cytometer (Beckman Coulter, Indianapolis, IN) using FlowJo software (Tree Star, Ashland, OR). For flow cytometric analysis, CD45⁺ cells were gated to exclude endothelial cells, hepatic stellate cells, and residue hepatocytes. Within CD45⁺ cells, CD44⁺ cells that bind to Chi3l1 were back-gated to determine the cells types.