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. 2009 Nov 18;35(3):792–805. doi: 10.1038/npp.2009.188

Figure 5.

Figure 5

AMI, but not VPA, reduces DNA methyltransferase activity of cortical astrocytes independent of DNMT-1 downregulation. (a, b) Drug treatment has no impact on DNMT-1 and GADD 45α expression levels. Cortical astroglial cells were treated with VPA, AMI, and TSA ranging from 24 to 72 h, including drug washout as specified. Western blot analysis was carried out using cellular extracts prepared from cortical astrocytes, which were probed with DNMT-1-, GADD 45α-, α-actin-, or hsp90-specific antibodies. (c) Cells were treated with VPA (10 mM), AMI (1,10 μM), and 5-Aza (8 μM) for 3d or 6–11d (5-Aza, long-term). Nuclear extracts were isolated and supplemented with the respective drugs. DNA methyltransferase activity was quantified by measuring the incorporation of S-adenosyl-L-[methyl-3H]methionine into the substrate Poly(dI-dC)·Poly(dI-dC). Data are presented as relative DNA methyltransferase activity±SE (n=2 for VPA/5-Aza, n=4 for AMI). Control is set to 100%.