Fig. 7
CCL9 promoted M2 polarization in macrophages via activating STAT6/ PPARβ pathway. A-C RAW264.7 cells was induced by 20 ng/mL IL-4, 20 ng/mL IL-13, 200 µM OA, 20 ng/mL CCL9 recombinant protein, and 500 nM J113863 (inhibitor of CCL9 receptor CCR1) for 48 h. A Immunofluorescence staining of CD206 in RAW264.7 cells. B mRNA expression of IL-10, Arg1 and YM1 in RAW264.7 cells were detected by real-time PCR method. C The protein levels of p-STAT6, STAT6, PPARβ and Arg1 in DEN/HFD induced liver tumors from MyD88 fl/fl and SMA MyD88−/− mice were detected by western blot analysis, and the gray value of p-STAT6, STAT6, PPARβ and Arg1 relative expression in livers of mice. PPARβ, Arg1 were normalized to β-actin, p-STAT6 was normalized to STAT6. D The protein levels of p-STAT6, STAT6, PPARβ and Arg1 in RAW264.7 cells were determined by western blot. The densities of proteins were quantified using densitometry. PPARβ, Arg1 were normalized to β-actin, p-STAT6 was normalized to STAT6. * p  < 0.05, ** p  < 0.01, *** p  < 0.001. (E-G) MyD88 fl/fl and SMA MyD88−/− mice were fed HFD for 3 months, then 1 × 10 6 Hepa1-6 cells were inoculated subcutaneously in these mice. The mice were fed HFD and blocked by intraperitoneal injection of CCL9 receptor CCR1 inhibitor J113863 (J113863, 3 mg/kg), and tumor growth were evaluated for 2 weeks. E Ex vivo images of resected tumors formed by subcutaneous injection of Hepa1-6 cells with or without CCR1 inhibitor (J113863) in MyD88 fl/fl and SMA MyD88−/− mice (n  = 4 per group). F Growth curves of tumor volume. G H&E staining, CD206 and CCL9 immunofluorescence staining and and CCL9 and MyD88 immunofluorescence double staining in transplanted tumors. Scale bar, 50 μm. H Statistical analysis. *** p  < 0.001