Figure 6.

Treatment of macrophages with SR141716A in vitro reduces the miR-466 family and miR-762 expression, promotes M2 polarization, and inhibits macrophage retention. Thioglycollate-elicited peritoneal macrophages were isolated from naïve mice and then cultured in 3T3-L1 adipocyte–conditioned medium containing SR141716A (10 μm) or DMSO for 24 h. A, qRT-PCR miRs. B–D, qRT-PCR of Klf4, Stat6, and Agap2. E, Netrin-1 ELISA from cell supernatants. For F and G, macrophages were stained with DilC₁₂(3) and treated with SR141716A or DMSO. The migration rate toward CCL19 (500 ng/ml) was assessed by seeding 2.5 × 10⁵ macrophages in serum-reduced conditioned medium from differentiated 3T3L-1 adipocytes in FluoroBlok plates. 12 h later, cell migration was quantified using Cytation5 imaging, and gene expression analysis was conducted for CCR7. F, the number of cells migrated toward CCL19. G, qRT-PCR expression of Ccr7. Data are shown as mean ± S.D. (error bars). **, p < 0.01; ****, p < 0.0001 versus DMSO vehicle control by two-tailed Student's t test.