Figure 2.

Ethanol induction of the Vamp2 gene requires transcriptionally activated HSF1. (A) HSF1 knock-down inhibits Vamp2 gene induction by ethanol. Overnight pretreatment of neurons with Hsf1 siRNA reduced the effects of 60 mM ethanol exposure for 1 h (E) on Vamp2 mRNA levels, while pretreatment with control siRNA had no effect [n ≥ 6; F_{(3, 44)} = 13.55; p < 0.001]. (B) Stimulation of the Vamp2 gene by ethanol is mediated by transcriptionally activated HSF1. Cortical neurons transfected with Hsf1-act showed increased Vamp2 mRNA expression, similar to the gene's induction by 60 mM ethanol for 1 h (E). Hsf1-inact transfection reduced ethanol induction of the Vamp2 gene, while Hsf1-inact transfection alone had no effect on basal Vamp2 mRNA levels. Control cultures were sham transfected with an empty pcDNA3.1+ construct [n ≥ 6; F_{(4, 73)} = 27.53; p < 0.001; ^*P < 0.05, ^**P < 0.01, ^***P < 0.001, or n.s. denotes no significance].