PMC

Adiponectin induces differentiation via Akt2. VSMC were transfected with siControl, siAkt-1 or siAkt-2 for 24 h, then treated with vehicle or 5 μg/ml adiponectin for 24h prior to western analysis. Arepresentative experiment is shown. Bar graphs were prepared from 3 separate experiments as in .

Adiponectin induces FoxO4 phosphorylation via Akt2. A, VSMC were treated with 5 μg/ml adiponectin for the indicated time points and harvested for western analysis. B, VSMC were transfected, treated, and analyzed as in . C, VSMC were transfected with plasmid encoding GFP or Flag-Foxo4 for 24h, then treated with vehicle or 5 μg/ml adiponectin for 24h and harvested for western analysis. For all panels, bar graphs were prepared from 3 independent experiments.

Adiponectin induces VSMC differentiation via S6K1 inhibition. VSMC were infected with control (GFP) or rapamycin-resistant mutant S6K1 adenovirus overnight, and then treated with vehicle or 5 μg/ml adiponectin for 24h prior to western analysis. Bar graphs were prepared from 3 separate experiments as in . p-values (Newman-Keuls multiple comparison post hoc tests) are indicated above the bars.

Adiponectin induces VSMC differentiation marker expression and modulates the mTORC1 signaling pathway. A, Human coronary artery VSMC were treated with recombinant human globular, trimeric or HMW-enriched adiponectin preparations as indicated and tested for 24 h prior to western blotting analysis with the primary antibodies indicated. B, VSMC were treated with 5 μg/ml HMW-enriched adiponectin for the indicated time points and harvested for western analysis as indicated.

Adiponectin and rapamycin have similar beneficial effects on vessel phenotype, but differ in adverse effects on endothelial cells. A, Freshly isolated pig femoral arteries were placed in organ culture for 3 days then treated with 100nM rapamycin or 10 μg/ml adiponectin for 4 days before harvesting for western analysis. Data from N=6 were quantitated with statistical analysis as in . B, HUVEC (top panels) or HUAEC (bottom panels) were treated with the indicated concentrations of rapamycin and adiponectin for 24h and harvested for western analysis. HUAEC blot is representative of N=3.

Adiponectin-induced VSMC differentiation requires AMPK. A, VSMC were pretreated with compound C (20 μM) for 30 min, followed by treatment with 5 μg/ml adiponectin for 24h and western blotting with the indicated antibodies. Bar graphs represent densitometric quantification (mean fold induction plus standard error of the mean) of 3 separate experiments. p-values (Newman-Keuls multiple comparison post hoc tests) are indicated above the bars. B, VSMC were transfected with siControl, siAMPKα1 or siAMPKα2 for 24h, then treated with vehicle or 5 μg/ml adiponectin for 24h and analyzed by western blotting. Data are representative of two experiments. C. VSMC were treated with 0.1 mM AICAR for the indicated time points and harvested for western blotting analysis as above. Data are representative of 3 separate experiments.