To directly test our model, we carried out a series of in vivo “single-cell genetic” analysis to compare phenotypes between DISC1 knockdown and multiple means of activating AKT signaling in newborn neurons, including overexpression of KIAA1212, overexpression of a constitutively active form of AKT1 (CA-AKT) (Kohn et al., 1996), and knockdown of PTEN. We used a previously characterized shRNA against Pten (PTEN-shRNA) (Jaworski et al., 2005) and confirmed its efficacy and specificity in knocking down endogenous PTEN in primary hippocampal neurons (Figure S1).
