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

Figure 2

Synergistic enhancement of precardiac gene expression by bone marrow MSCs in response to coaddition of TSA with BIX01294. (a) MSCs were incubated for 48 hrs with various doses of BIX01294 in absence or presence of 50 nM TSA, prior to harvesting RNA and measuring Mesp1 expression by qPCR. On the left side of the chart, the y-axis is normalized to Mesp1 levels obtained from MSC cultures treated with 8 μM BIX01294. The right side y-axis is scaled to the relative increase in Mesp1 expression resulting from the TSA coaddition to a given concentration of BIX01294. For this series of experiments, BIX01294 generated an ~160-fold increase in Mesp1 expression as compared to control levels, with TSA coaddition enhancing further the Mesp1 response an additional 5.6-fold. The numbers of biological repeats represented in the graphed data are as follows: 2 μM BIX01294 and 2 μM BIX01294+TSA (), with remaining groups having values = 14. (b) RNA was assayed for brachyury expression by qPCR, following its isolation from MSC cultures incubated for 48 hrs in the absence or presence of 8 μM BIX01294 and/or 50 nM TSA. The coaddition of TSA synergistically enhanced BIX01294-stimulation of brachyury expression 7.2-fold (; ; ).

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