For α5 subunit re-expression studies, the mouse α5 nAChR subunit gene,Chrna5, was cloned into the pCDF1 lentivirus expression vector containing cop-GFP from Systems Biosciences, Inc. (Mountain View, CA). For α5 subunit knockdown studies, two different short hairpin interfering RNAs (shRNA) directed against the rat Chrna5 gene were were designed using the Genscript, Inc. online construct builder (see Supplementary Figs for shRNA sequence). The shRNAs were cloned into the pRNAT-U6.2/Lenti construct containing GFP (GenScript, Piscataway, NJ). Control vectors were identical to the expression constructs, but without the gene insert.
