PKB phosphorylation in cerebellar slices.

Cerebellar slices from WT mice were treated with 1 µM wortmannin or vehicle control (DMSO) for 10 min or with API-2 for 30 min in their recovery chamber before addition of 10 µM DEA/NO for another 10 min. Total lysates were prepared and subjected to Western blotting with phospho-S473 and phospho-T308 specific PKB antibodies as well as total PKB antibodies as described in Materials and Methods. Blots shown are from one experiment representative of three. Bar graphs show densitometric analysis of PKB phosphorylation on T308 or S473, as indicated, compared to levels in untreated “control” samples and are mean ± s.e.m. of three experiments.