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Fig. 2 PKA modulators influence adipogenesis, gene expression for adipogenesis, osteogenesis and osteoclast-inducing factors in primary MSCs. (A) Enhancement of adipogenesis by IBMX and forskolin. Cells were treated in complete growth medium [C] or in adipocyte induction medium without [A] or with the indicated modulator of PKA activity [I: IBMX (0.45 mM), F: forskolin (100 μM), P: PKI (20 μM), I+P: IBMX+PKI, F+P: forskolin+PKI] for 7 and 14 days. Differentiated adipocytes were detected by the accumulation of lipids, which were stained with Oil Red O. (B) Oil Red O staining as OD510. The results displayed in the bar graphs are quantitative analytic data from three independent experiments and are presented as the meanSD values. Each ratio was normalized to the control, and significance was determined by Student's t-test. (** p<0.01 versus C; # p<0.05, # #<0.01 versus A, # # # p<0.05 versus I or F). Real-Time RT-PCR analysis of the gene expression ratio of PPARγ2/GADPH (C), Runx2/GADPH (D), and RANKL/OPG (E) after treatment with PKA modulators for 7 days.