Fig 3. PIC Ⅱ enhances the function of M1-polarized macrophages and promotes the release of chemokine CXCL16. (A) The heatmap and clusters of M1 type-related macrophage markers. (B) GSEA analysis of M1-polarized macrophage activation. Relative mRNA levels of (C) Il6 ,Inos , Cd163 , Tgm2 , (D) Mmp13 , Mmp2 ,Fn1 and Cxcl16 in mice liver tissue were measured by qPCR and further normalized with Hprt1 . (E) The protein levels of CXCL16 were measured by western blot and normalized by β-ACTIN in the mouse liver. Relative mRNA levels of (G) Cxcl16 in macrophages were measured by qPCR and further normalized with Hprt1 . (H) The protein levels of CXCL16 in macrophages and derived CM were measured by western blot and normalized by β-ACTIN. Representative images of immunofluorescence staining for F4/80 and CXCL16 (F) in the liver (Scale bar = 20 μm) and (I) macrophages (Scale bar = 100 μm). Statistical significance: *P < 0.05, **P < 0.01, ***P < 0.001, compared with the control group;#P < 0.05,##P < 0.01,###P < 0.001, compared with the model group (n = 6).