Nanostring of gene expression in intrahepatic leukocytes from wild type (WT) or myeloid-specific RAGE knockout (RAGE-MKO) mice fed a chow or high-fat, -fructose, and -cholesterol (FFC) diet

Intrahepatic macrophages in nonalcoholic steatohepatitis (NASH) are heterogenous and include proinflammatory recruited monocyte derived macrophages. RAGE is expressed on macrophages and can be activated by damage associated molecular patterns (DAMPs) upregulated in NASH, yet the role of macrophage-specific RAGE signaling in NASH is unclear. Therefore, we hypothesized that a subset of RAGE expressing macrophages are proinflammatory and mediate liver inflammation in NASH. We profiled the transcriptome of intrahepatic leukocytes in a murine model of diet induced NASH in WT and RAGE-MKO mice. By Nanostring nCounter Immunology Codeset Analysis of IHLs, we observed that myeloid-specific RAGE deletion resulted in amelioration of activation of macrophage and T cell pathways which was associated with attenuation of liver inflammation. C57BL/6J WT and RAGE-MKO mice were fed a high-fat, -fructose, and -cholesterol (FFC) diet to induce NASH for 6 months. After enzymatic digestion of equal amounts of liver tissue, IHLs were isolated by density Percoll-gradient ultracentrifugation. Gene expression analysis was conducted using nCounter Mouse Immunology Codeset (Nanostring, Seattle, WA) on IHLs from 4 biological replicates within each group.