Figure 1.

Adiponectin Stimulates Exosome Production by hMSCs through T-cadherin

Exosomes were isolated from the culture medium of hMSCs as described in Materials and Methods. Exosome and whole-cell lysate (WCL) were lysed after 48 h with or without high-molecular-weight adiponectin (HMW-APN, 20 μg/mL). (A) NTA (n = 3). (B) Western blot analysis with the indicated antibodies of typical exosome markers, adiponectin (APN), and T-cadherin (T-cad), as indicated (n = 3). (C) hMSCs were treated with HMW-APN for 48 h in a dose-dependent manner, and the isolated exosomes were subjected to western blot analysis using the indicated antibodies (n = 2–3). (D) qPCR analysis of control and T-cad (CDH13) RNAi-transfected hMSCs at 48 h after transfection (n = 2). (E) Exosomes and WCL were collected from T-cad RNAi-transfected cells with or without APN. The proteins were subjected to western blot analysis with the indicated antibodies. Representative immunoblots are shown. Data are mean ± SEM. The results of the experiment were tested in two separate trials. For (A) and (B), Student’s t test; for (C), one-way analysis of variance with Dunnett’s multiple comparisons. ∗p < 0.05; ∗∗p < 0.01; ∗∗∗p < 0.001.