Inhibition of G9a Histone Methyltransferase Converts Bone Marrow Mesenchymal Stem Cells to Cardiac Competent Progenitors

Figure 2

MSCs express precardiac markers in response to BIX01294. (a) MSCs were exposed to multiple compounds over a range of concentrations, with RNA harvested and analyzed by qPCR. Chart summarizes results with optimized doses of inhibitors specific for G9a HMTase (BIX01294), nitric oxide synthase (3-bromo-7-nitroindazole; BNI), GSK3β (CHIR99021), BMP (dorsomorphin), Wnt (IWP4), TGFβ (1,5-naphthyridine pyrazole derivative-19; NPy19), FGF (PD173074), β-catenin (PNU74654) c-Jun N-terminal kinase (SP600125), and histone deacetylase (trichostatin A; TSA). Only BIX01294 was able to significantly enhance Mesp1 and brachyury expression over control levels. Note that the relative levels of gene expression are shown in log scale. (b) Parallel cultures of MSCs treated for 48 hrs with various concentrations of BIX01294 and assayed for Mesp1 gene expression using real-time qPCR. MSCs exhibited the highest levels of Mesp1 expression when exposed to 8 μM BIX01294. (c) MSCs exposed to 8 μM BIX01294 for up to 8 days and examined for Mesp1 gene expression, with optimal responses obtained at 48-hour incubation. ; ; .

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