Table 2. VPA/TSA and AMI Trigger CCpGG DNA Hypomethylation.
| Treatment |
Change in DNA methylation analyzed by LUMA |
|||||
|---|---|---|---|---|---|---|
| Drug |
24 h |
72 h |
72 h+48 h washout |
|||
| CTX | HPC | CTX | HPC | CTX | HPC | |
| 1 mM VPA | −4.2±3.3 | −9.7±2.4* | −4.9±1.8* | −5.2±0.6* | −0.6±2.8 | −6.7±2.4 |
| 10 mM VPA | −7.5±1.3* | −4.0±2.7 | −17.2±2.0* | −5.0±1.4* | −2.6±4.1 | −6.0±3.1 |
| 100 μM LTG | n.d. | n.d. | +0.5±3.0 | n.d. | n.d. | n.d. |
| 1 μM AMI | −7.4±4.2* | n.d. | −9.7±4.0* | n.d. | −4.4±1.3 | n.d. |
| 10 μM AMI | −4.3±2.8 | n.d. | −9.2±2.0* | n.d. | −6.5±3.5 | n.d. |
| 0.2 μM TSA | −7.1±3.7* | −9.6±1.0* | −11.6±5.0* | −10.4±2.3* | n.d. | n.d. |
Abbreviation: n.d., not determined.
Primary astrocytes were treated with different concentrations of VPA, LTG, AMI, and TSA as specified in the table. Levels of DNA methylation were measured after 24 and 72 h, as well as after the washout period. For all experiments, genomic DNA was isolated from the cells and 2 μg was subjected to luminometric methylation analysis (LUMA). The results are expressed as mean±SE of the percentage of the change in CCpGG DNA methylation compared with untreated cells. The experiments were performed in two to four independent trials, each with six technical replicates; t-test: *p<0.05 vs untreated cells.