Figure 2.
Primary cultures of cortical neurons derived from PDK1 wild type (WT) and PDK1 K465E knock-in (KI) E15,5 embryo littermates were generated as described.27 After 6 d in culture, cells were incubated in complete medium or deprived of serum for 24 h in the absence or presence of 50 ng/ml of Brain-Derived Neurotrophic Factor (BDNF) and the indicated concentrations of the AZD8055 inhibitor. Cell viability was determined by the MTT (1-(4,5-Dimethylthiazol-2-yl)-3,5-diphenylformazan) reduction assay and is represented as the mean ± standard errors of the mean from 5 independent embryos per genotype, with each sample assayed in quadriplicate. As a control for the different treatments, cell lysates from matched PDK1 wild type and PDK1 mutant littermate mice were immunoblotted with the indicated antibodies, where the first 8 lanes and the last 2 lanes were generated from separate gels. **, P < 0.01 between genotypes as determined by the Student's t test.