Inhibition of Histone Methyltransferase, Histone Deacetylase, and β-Catenin Synergistically Enhance the Cardiac Potential of Bone Marrow Cells

Figure 1

TSA synergistically enhances stimulation of Mesp1 expression by BIX01294. MSCs were incubated with various compounds plus or minus BIX01294 over a broad range of concentrations, with RNA harvested 2 days later and analyzed for Mesp1 expression by qPCR. The chart presents summarized results using optimized doses of inhibitors specific for G9a HMTase (BIX01294), DNA methylation (5-azacytidine (5aza)), nitric oxide synthase (3-bromo-7-nitroindazole (BNI)), GSK3β (CHIR99021 (CHIR)), Wnt (IWP4), TGFβ (1,5-naphthyridine pyrazole derivative-19 (Npy19)), and histone deacetylase (trichostatin A (TSA)). The numbers of biological repeats represented in the graphed data are as follows: control and BIX01294 () and BIX01294+TSA (), with remaining groups having values ranging from 3 to 5. BIX01294 was the only reagent by itself that was able to induce Mesp1 expression, with >83-fold increase over control levels (). None of the other drugs were able to increase Mesp1 expression. However, of these latter drugs, only TSA when combined with BIX01294 produced a significant enhancement of Mesp1-expression over the levels obtained from the BIX01294 only treated cultures (). Note that the relative levels of Mesp1 gene expression are normalized to levels obtained from BIX01294-treated MSC cultures.