Fig. 4

Roles of macrophage-derived MVs in insulin-stimulated pAkt levels in human differentiated adipocytes. a. Human differentiated adipocytes were fixed and stained with oil red O for lipid accumulation. (200×). b. The optical absorbance of lipid accumulation was measured at 510 nm. c. The mRNA levels of PPARγ1 and PPARγ2 in preadipocytes and adipocytes were analyzed by qRT-PCR. d. Protein levels of pAkt in human differentiated adipocytes were detected by western blotting. Adipocytes were pretreated with various reagents for 24 h, and then the cells were stimulated with 0, 1, 10, or 100 nM insulin for 20 min, lysed with lysis buffer, and prepared for the immunoblotting assay. Total Akt (tAkt) was used as a loading control. Western blot images were analyzed using Bandscan software, and statistical analysis is presented below. *P < 0.05, **P < 0.01 and ***P < 0.001 indicate a significant up-regulation compared with the corresponding vehicle groups. #P < 0.05 and ###P < 0.001 indicate significant difference when compared with indicated conditions