Elevation of pAkt levels in primary neurons treated with Aβ oligomers and in neurons from the R1.40 transgenic mouse model of AD. A. Cultured primary cortical neurons (21 DIV) were treated with vehicle (Veh), 2.0 μg/ml Aβ monomers (AβM) or 2.0 μg/ml Aβ oligomers (AβO) for 24 hours. Western blot analysis of cell extracts with antibodies against pAktS473, total Akt, p4E-BP1S65 and GAPDH revealed a slight increase in the levels of pAkt and p4E-BP1 in AβO treated cultures when compared to vehicle or AβM treated cultures. B-C. Quantification of the ratios of pAkt/total Akt (B) and p4E-BP1/GAPDH (C) revealed statistically significant increases upon exposure to AβO (2.0 μg/ml) when compared to AβM treatment (2.0 μg/ml) or vehicle controls (p < 0.05 for AβO versus vehicle or AβM; unpaired t test; mean ± SEM; n = 3 independent treatments). D. Western blot analysis of detergent soluble cell extracts of from cultured primary cortical neurons (21 DIV) from non-transgenic (WT) and homozygous R1.40 (R/R) mice with antibodies against pAkt (pAktS473), p-mTOR (p-mTORS2448) and GAPDH revealed elevated levels of pAkt (pAktS473) but not p-mTOR in the R/R neurons when compared to WT neurons. E-F Quantification of the ratios of pAkt/GAPDH (E) and p-mTOR/GAPDH (F) revealed statistically significant increases in the levels of pAkt/GAPDH (p = 0.04; unpaired t test; mean ± SEM; n = 3 independent cultures) but not p-mTOR/GAPDH (p = 0.74; unpaired t test; mean ± SEM; n = 3 independent cultures) in cell lysates form the R/R cultures.