Fig. 4.
P2X4R antagonist selectively decreases alcohol-induced transcription activity. (A) Alcohol promotes transcriptional activity of CREB. ESdM cells (6 × 105) were grown for 4 days to confluence and treated with alcohol (100 mM) for 2 h. Nuclear protein extracts were harvested CREB activity were measured by TransAM™ CREB protein assay. Microglial cells treated with alcohol for 2 h showed a statistically significant increase in CREB (*P < 0.001) when compared with control. Data are represented as fold change compared with untreated control. Data were analyzed using a two-tailed t-test. Data points represent mean ± SEM (n = 3). (B) Treatment with 5-BDBD significantly blocked CREB activity compared with alcohol treatment alone (***P < 0.0001). Data are represented as fold change compared with control. Results were determined using a two-way ANOVA with Bonferroni post hoc for multiple comparisons. Data points represent mean ± SEM (n = 3).