Figure 6.

Demethylation of BDNF promoter I and increased BDNF mRNA expression occurs after DNMT inhibition and both are dependent on synaptic activity. A, Bisulfite modification of genomic DNA followed by quantitative PCR (Q-PCR) to measure levels of unmethylated BNDF promoter I. Representative gel electrophoresis of Q-PCR results after control, PTX, and 5azaC treatments with and without TTX or AP-5. B, Bar graph shows a significant increase in the level of unmethylated BDNF promoter I after 5azaC and PTX treatments compared with controls (black and gray bar indicates mean ± SEM) but no changes in the level of unmethylation when 5azaC treatments were in combination with TTX or AP-5 (*p < 0.05; **p < 0.01). C, Bar graph showing increased BDNF mRNA in response to both 5azaC and PTX treatments and decreased expression with 5azaC plus TTX or AP-5 cotreatments (*p < 0.05; **p < 0.01; ***p < 0.001). The numbers on the bars indicate the number of experiments (B, C).