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Fig S1. Effects of Akti1/2 (40 µM) on mdMSC differentiation. A. Added to adipogenic medium Akti1/2 enhanced expression of adipogenic genes aP2 and PPARγ at 3 days. B. Akti1/2 decreased expression of osteogenic genes osterix and RUNX2, assayed by qRT-PCR and, shown in C, decreased activity of alkaline phosphatase (ALP).

Fig S2. Effects of KU63794 (2 µM) on mdMSC differentiation. A. Added to adipogenic medium, KU63794 caused mdMSC cell death at concentrations used in short term studies, shown by the XTT assay. B. mdMSC were more resistant to mTOR inhibition when cultured in the osteogenic medium, allowing assay of alkaline phosphatase (ALP). KU63794 decreased ALP activity at 4 days.

Table S1. Insulin (100 ng/ml) alone does not induce mdMSC differentiation. Insulin was added to mdMSC in growth medium, and RT-PCR for adipogenic genes measured at 4 days (A), and osteogenic genes at 7 days (B), similar to times assayed in Figure 7. Gene expression was compared to that found in typical adipogenic and osteogenic medium, which caused significant changes in respective genes. Insulin did not induce measurable differentiation.

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