Fig. (4). Nicotinamide (NIC) induces phosphorylation of Bad during anoxia exposure.
Equal amounts of neuronal protein extracts (50 μg/lane) were immunoblotted with anti - phospho-Bad (p-Bad, Ser136) antibody. The expression of p-Bad was increased 2-6 h following the exposure to a 4 h period of anoxia (A). NIC (12.5 mM) applied 1 h prior to anoxia further increased the phosphorylation of Bad during anoxia over a 24 h period (B). Application of the Akt1 phosphorylation inhibitor wortmannin (W, 500 nM) or LY294002 (LY, 10 μM) 1 h prior to anoxia was sufficient to block the expression of p-Bad in the presence of NIC 3 h later (C). Con=Control.