Band densities for all samples on a given gel were normalized to the band thickness for a sample from a dog subjected to sham operation, treated with vehicle and killed 1 h after surgery, allow comparisons from experiment to experiment. Each gel included at least one sample from such a control animal allow comparisons of data across different studies, and a control animal was Doxorubicin Topoisomerase inhibitor prepared for each experiment. These antibodies were found in this study: 1) antiphospho Akt rabbit polyclonal antibody, which recognizes Akt only when phosphorylated at Ser473, 2) anti Akt rabbit polyclonal antibody, which recognizes total Akt, 3) anti phospho GSK 3B rabbit polyclonal antibody, which recognizes GSK 3B phosphorylated at Ser9, 4) anti GSK 3B mouse monoclonal antibody, which recognizes total GSK 3B, 5) antiphospho FOXO3A rabbit polyclonal antibody, which recognizes FOXO3A phosphorylated at Ser253, 6) anti FOXO3A rabbit polyclonal antibody, which recognizes total FOXO3A, 7) anti phospho MAPK mouse monoclonal antibody, which recognizes ERK1 and ERK2 phosphorylated on both at Thr202 and Tyr204 remains, 8) anti MAPK1/2 rabbit polyclonal antibody, which recognizes total ERK1/ERK2, 9) anti B actin mouse monoclonal antibody, which recognizes an positioned within the N terminal domain of the B isoform of actin. Secondary antibodies for Westerns were horseradish peroxidaseconjugated Mitochondrion donkey anti rabbit IgG for polyclonal antibodies, or sheep anti mouse IgG for monoclonal antibodies. Caspase activity assays were performed on fresh frozen brain sections using the APO LOGIXTM carboxy fluorescein caspase detection set according to the manufacturers instructions. FAM DEVDFMK is really a carboxy fluorescein marked analog of zDEVD fluoromethyl ketone, a wide range cysteine protease inhibitor that binds activated caspases enters cells and irreversibly. FAM DEVD FMK exhibits higher affinity for caspase3 than for caspase 8, caspase 7, caspase 10 or caspase 6 and exhibits much lower affinity for the calpains than for caspases, hence, at 5 uM FAM DEVDFMK is really a somewhat selective inhibitor of caspase 3. Moreover, FAM DEVD FMK labeling of Hesperidin molecular weight CA1 neurons fits well with caspase 3 activation, as assessed by Western blot analysis. In this study we thus make reference to FAM DEVD FMK labeling as indicative of caspase 3 activity. In temporary, estradiol and vehicleinjected animals were deeply anesthetized with pentobarbital and killed by decapitation at 2-4 h after ischemia or sham operation. Brains were removed, frozen and cut into sections in the coronal plane of the dorsal hippocampus. Brain sections were labeled with 5 uM FAM DEVD FMK, washed three times with 1 Working Dilution Wash Buffer and considered under a ECLIPSE TE 300 fluorescent microscope equipped with an image analysis program at an wavelength of 488 nm and emission wavelength of 565 nm.