Fig. 5

Adiponectin enhanced osteoblast differentiation via the AMPK pathway. A Id promoter site was fused to luciferase and transfected into MC3T3-E1 cells using Lipofectamine 3000. The cells were treated with AdipoRon (50 μM) with or without AICAR (500 μM) for the indicated times during culture, and luciferase activity was assessed as a measure of Id transcriptional activity. The data are the mean from four culture wells (mean ± SEM). **Indicates P < 0.01 vs. treatment with AdipoRon alone. B MC3T3-E1 cells were incubated with AdipoRon. Expression of the osteoblast differentiation-related genes Runx2, Osx, and Ocn in MC3T3-E1 cells after incubation with AdipoRon. The samples were analyzed by quantitative reverse transcription polymerase chain reaction and were normalized to the ß-actin mRNA. The data are the mean from six culture wells (mean ± SEM). **Indicates P < 0.01 vs. treatment with AdipoRon alone. C Cells were treated with AdipoRon in the presence or absence of AICAR (500 μM). Western blotting was conducted using targeted and β-actin antibodies. Similar results were obtained in four independent experiments