Two weeks following the conclusion of behavioral studies (~9 weeks after stereotactic injections) Clic4 expression from AAV-CLIC4 injected animals was detected in vivo by immunohistochemistry using antisera against the FLAG epitope tag. Ethanol regulation of endogenous Clic4 mRNA in mouse brain was determined through quantitative real-time PCR (qRT-PCR) to confirm prior microarray studies (Kerns et al., 2005). Additional details are provided as Supplementary Material.
