FIGURE 2

Preconditioning ischemia activates CREB in the cortex. (A) Western blotting showing expression of Bcl-2, CREB, phosphorylated CREB (Ser 133) and CBP after preconditioning 30-minute MCAO. Images shown are representative blots from three independent experiments using pooled tissue from two animals. (B) Immunohistochemistry showing CREB, phosphorylated CREB and CBP expression after 30-minute MCAO in the region of the cortex made tolerant by preconditioning ischemia, compared with control (sham-operated animals). The schematic brain to the right shows the region of brain made tolerant by preconditioning in gray, and the region studied in red. Images shown are 40 and representative of two independent experiments, scale bar represents 50 m. (C) Increased interaction of CREB with bcl-2 CRE after 30-minute preconditioning. After preconditioning, nuclear extracts are incubated with a labeled bcl-2 CRE oligonucleotide and separated by electrophoresis. Note the increased retention of oligonucleotide probe by nuclear extracts. The interaction was reduced when nuclear extracts were incubated with excess cold oligonucleotide (right lane). Data shown are a representative blot of two experiments.