Figure 8. CDDO-Me reduces CCL3-CCR1 and CCL4-CCR5 expression in macrophages
(a) The top pathway in Regulatory Effects detected by Ingenuity Pathway Analysis using 195 genes downregulated in Figure. 5a. (b) Heatmap shows the gene expressions involved in chemotaxis in the liver. (c and d) The serum protein levels of CCL3 (c) and CCL4 (d) measured using ELISA (mean + S.D., n = 8). Statistical differences were analysed using Dunnett’s multiple comparison test (*P < 0.05; **P < 0.01; ***P < 0.001, ****P < 0.0001 versus vehicle-treated CDAHFD-fed mice). (e) Hepatic sections stained with anti-CCR1 or anti-CCR5 antibody. Scale bars represent 50 μm. (f and g) CCR1 (f) or CCR5 (g)-positive cells quantified in the hepatic sections from Figure. 8e (mean + S.D., quadruplicate). (h) Sections from CDAHFD-fed mice multiple stained with anti-CCR1(green), anti-CCR2 (red), anti-Ly6G (blue), and anti-CLEC4F antibodies (pink) (left panel) or anti-CCR5 (green), anti-CCR2 (red), anti-GPNMB (blue), and anti-CLEC4F antibodies (pink) (right panel). Scale bars represent 50 μm. (i-l) mRNA expression of Ccl3 (i), Ccl4 (k) and the receptors, Ccr1 (j) and Ccr5 (l) determined using quantitative PCR in RAW264.7 cells treated with CDDO-Me for 6 h (mean + S.D., triplicate). Cont, Control; Veh, Vehicle.