Effects of siRNA-AdipoR1 transfection or an AICAR treatment on Runx-2 mRNA and protein expressions in MC3T3-E1 cells. (A and B) Total RNA was collected at 4, 7, 10 days after siRNA-AdipoR1 transfection, and cell lysate was collected at 7 days after transfection. Real-time PCR and immunoprecipitation were performed as described in MATERIALS AND METHODS. Neither Runx-2 mRNA (A) nor its protein (B) expressions were changed by blocking the receptor expression. (C and D) AICAR (0.5 mM), an AMP kinase stimulator, was added after the cells reached confluency. Total RNA was collected at 14, 21, 28 days, and cell lysate was collected at 14 days. Neither Runx-2 mRNA (C) nor its protein (D) expressions were changed by AICAR. The result was the representative of five different experiments.