3% H2O2, and non specific binding was prevented by incubation in PBS 10% Goat Serum. The first antibody was diluted 1,25 in PBS 10% Goat kinase inhibitor Wortmannin Serum, and the slides were incubated overnight at 4 C in humidified chamber. Slides were then washed in 1% Triton PBS, following a second blocking step with PBS 10% Goat Serum. Secondary bioti nylated antibody was added at the dilution of 1,100. After 30 minutes of incubation at room temperature, sections were washed in PBS 1% Triton X 100 followed by only PBS, before staining by use of a Vectastain peroxidise standard ABC kit, and DAB solu tion, both according to the suppliers protocol. Cells were then counterstained with haematoxylin and mounted with glycerol solution. For each sample, six random fields at 40 �� of magnification were analyzed by counting positive cells with respect to total cells.

Results were expressed as an immunohistochemical score, based on the German Immunoreactive Inhibitors,Modulators,Libraries Score, which combines quantity and intensity values. The IHS is calculated by combining the Inhibitors,Modulators,Libraries percentage of immunoreactive cells with an estimate of the staining intensity, quantity score, no staining is scored as 0, 1 to 10% of cells stained scored as 1, 11 to 50% as 2, 51 to 80% as 3, and 81 to 100% as 4, staining intensity was rated on a scale of from 0 to 3, with 0 being negative, 1 weak, 2 moderate, and 3 strong. The raw data were converted to IHS by multiplying the quan tity and staining intensive score. Apoptosis Apoptotic nuclei in paraffined tissue were identified by Terminal deoxynucleotidyl Transferase Biotin dUTP nick end labeling technique.

The procedure was the one reported in the manufacturers instruction. Apoptotic nuclei were identified by the red precipitate obtained by incubating the glass slides with 0. 04% 3 amino 9 ethyl carbazole in 50 mM sodium citrate buffer, pH 5 containing 0. 015% H2O2. The values were expressed as the percen tage Inhibitors,Modulators,Libraries of TUNEL position nuclei with respect to the total haematoxylin Inhibitors,Modulators,Libraries stained nuclei. Six different fields with about 40 total cells for each sample were measured. Study protocol in vitro Cell culture b glycero phosphate, 0. 1 mM L ascorbic 2 phosphate and dexametasone were purchased from StemCell. Almost all other chemicals were from Sigma. Other companies are specified in the text. Bone marrow cells were obtained from 8 to 10 week old Sprague Dawley rats.

The femurs were removed aseptically and dissected, the ends of bones were cut and the marrow was flushed out with DMEM by using a needle and syringe. The marrow was disperded gently by pipetting several Inhibitors,Modulators,Libraries times KOS 953 to obtain a single cell suspension and the cells were counted with a hemocytometer. Cells were seeded into 25 ml cell culture flasks at a density of 1 �� 106 cells ml and culture for 48 h at 37 C in a humi dified atmosphere with 5% CO2 in DMEM containing 2 mM L glutamine, 20% fetal bovine serum, 100 U ml streptomycin and 100 ug ml penicillin.