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. 2017 Jul 16;2017:3464953. doi: 10.1155/2017/3464953

Figure 1.

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 (n = 31) and BIX01294+TSA (n = 18), with remaining groups having n 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 (∗∗p < 0.005). 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 (∗∗p < 0.005). Note that the relative levels of Mesp1 gene expression are normalized to levels obtained from BIX01294-treated MSC cultures.